Teaching AS Biology

Practical Skills

University of Cambridge International Examinations

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Tel: +44 1223 553554 Fax: +44 1223 553558 E-mail: [email protected] Website: www.cie.org.uk

© University of Cambridge International Examinations 2005

PSAS9700 0501
 Teaching AS Biology Practical Skills
in preparation for 9700 papers 31 and 32

Your attention is drawn to the section on Risk Assessment on page 17 of the

Introduction to this booklet, and to the hazards indicated in Appendices 1 and 2.
While all effort has been made to ensure that appropriate safety indications are
given, CIE accepts no responsibility for the safety of these experiments and it is
the responsibility of the teacher to carry out a full risk assessment for each
experiment undertaken, in accordance with local rules and regulations. Hazard
data sheets should be available from your suppliers.
Contents

Introduction 1
Why should I read this booklet? 1
How much teaching time should I allocate to practical work? 1
Can I use the practicals in these booklets in a different order? 1
What resources will I need? 2
Is there a limit to the class size? 2
Why should I teach my students practical skills? 2
Points to consider 2
What are the practical skills required by this course? 4
Summary of each of the seven skills 5
Ways of doing practical work 9
Keeping records 12
How is a practical activity organised? 13
Risk assessment 15
Eye protection 17
AS Skills 18
Teaching students to manipulate, measure and observe 18
Teaching students to present data and record observations 18
Teaching students to analyse, draw conclusions and evaluate 19
Appendix 1 - Designing a practical course for AS 20
Syllabus section A – Cell Structure 21
Syllabus section B – Biological Molecules 21
Syllabus section C – Enzymes 22
Syllabus section D – Cell Membranes and Transport 23
Syllabus section E – Cell and nuclear division 24
Syllabus section F – Genetic Control 24
Syllabus section G – Transport 24
Syllabus section H – Gas Exchange 25
Syllabus Section I – Infectious disease 26
Syllabus Section J – Immunity 26
Syllabus Section K – Ecology 26
Appendix 2 – practicals for which full details are provided 27
Practical 1 - Investigation into size and scale of microscopic tissues 27
Practical 2 - Microscopic observation of cells and tissues 33
Practical 3 - The identification of biological chemicals present in solutions 40
Practical 4 - Investigation of the carbohydrates metabolised by yeast 45
Practical 5 - The effect of pH on enzymes 51
Practical 6 - The effect of inhibitors on enzyme activity 57
Practical 7 - The effect of temperature on membrane permeability in beetroot 62
Practical 8 - Broad bean root tip squash 67
Practical 9 - The extraction of DNA from onions 73
Practical 10 - The effect of wind speed on the rate of transpiration in a leafy shoot 78
Practical 11 - Investigating the role of carbon dioxide in living organisms. 86
Introduction
You may have been teaching AS and A level biology for many years or perhaps you
are new to the game. Whatever the case may be, you will be keen to ensure that you
prepare your students as effectively as possible for their examinations. The use of a
well-structured scheme of practical work will certainly help in this ambition. However it
can do so much more. Scientists who are thoroughly trained and experienced in
practical skills, will have a ‘feel’ for the subject and a confidence in their own abilities
that is far greater above those with a purely theoretical background. It is true that there
are branches of biology that might be described as purely theoretical but they are in the
minority. Essentially, biology is a practical subject and we owe it to our students to
ensure that those who pursue science further have the necessary basic practical skills
to take forward into their future careers. Furthermore, the basic skills of planning,
analysis and evaluation will be of great value to those who pursue non-science careers.

Why should I read this booklet?

Some of you may be wondering why you should need a booklet like this. If your
practical skills are of a high order and you feel confident teaching these skills to others,
you probably don’t need it; but you might find some of the exercises described in the
appendices useful. However, if you are like the majority of us, a little help and support
is likely to be appreciated. This booklet aims to provide at least some of this support.
It is designed for the teacher rather than for the student. Its objective is to provide a
framework within which the practical skills of teachers can develop and grow.
Experience shows that as a teacher’s practical skills grow, so too do the confidence to
teach such skills and the time that you will be prepared to spend on teaching practical
work.

How much teaching time should I allocate to practical work?

The syllabus stipulates that at least 20% of teaching time should be allocated to
practical work. This is in addition to any time the teacher chooses to use for practical
demonstrations to illustrate the theory syllabus. This emphasis on practical work is not
misplaced. Consider the weighting given to assessment objectives in the syllabus: 24%
is allocated to experimental skills and investigations and 30% is allocated to handling,
applying and evaluating information. Taken together, 55% of the total award is related
to a students’ ability to interpret data, understand how this has been obtained,
recognise limitations and suggest explanations; all of which lend themselves to
investigative work involving practical experience. If the specific practical papers are
considered in isolation, they still represent 23% of the AS and 24% of the A Level
award.
In planning a curriculum, teachers should therefore expect to build in time for
developing practical skills. If, for example, the time allowed is 5 hours per week over 35
weeks, then a minimum of 1 hour per week should be built into the plan, so that over
the year, a minimum of 35 hours is made available. Bearing in mind the emphasis on
assessment objectives that related to information handling and problem solving, a
minimum of 2 hours per week might be more appropriate, which at 40% of the time is
still less than the overall weighting for these assessment objectives.

Can I use the practicals in these booklets in a different order?

It is assumed in these booklets that for A level candidates, the AS work will be taught in
the first year of the course, with the A2 work being covered in the second year. If the
linear A Level assessment route is used, care should be taken with regard to in the

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order in which practical exercises are used, as the skills practiced in these booklet are
hierarchical in nature, i.e. the basic skills established in the AS booklet are extended
and developed in the A2 Level booklet. Thus, students will need to have practiced
basic skills using AS exercises before using these skills to tackle more demanding A
Level exercises.
The exercises in these booklets are given in syllabus order. A teacher may well decide
to use a different teaching sequence, but the point made above, regarding AS/A2
exercises, still applies.

What resources will I need?

For a practical course in A-level Biology to be successful, it is not necessary to provide
sophisticated equipment. Some of the more advanced practicals in these booklets may
require less easily obtainable equipment, but the vast majority can be performed using
the basic equipment and materials in the lab. Alternative ‘low-tech’ exercises are also
provided where possible.
A list of the basic resources required for assessment may be found in the syllabus. A
more detailed list may be found in the booklet ‘CIE Planning For Practical Science in
Secondary Schools’, Appendix B.

Is there a limit to the class size?

It is true that there is a limit to the class size that is manageable in a laboratory
situation, particularly when students may be moving about. The actual size may be
determined by the size of the room, but as a general guide, 15 - 20 students is the
maximum that one person can reasonably manage, both for safety reasons and so that
adequate support can be given to each student. Larger numbers can more easily and
safely be accommodated with input from another person with appropriate qualifications
/ experience or splitting the class into two groups for practical lessons.
Why should I teach my students practical skills?
Although this section is likely to be read once only, it is arguably the most important;
for, if it convinces readers that practical work is an essential part of biology as a
science and underpins the whole teaching programme, the aim of publishing this
booklet will have been achieved.

Points to consider
• It’s fun! The majority of students thoroughly enjoy practical work. The passion that
many scientists have for their subject grew out of their experiences in the practical
classes. Students who enjoy what they are doing are likely to carry this enthusiasm
with them and so be better motivated.
• Learning is enhanced by participation as students tend to remember activities they
have performed more easily, thus benefiting their long-term understanding of the
subject. Students who simply memorise and recall facts find it difficult to apply their
knowledge to an unfamiliar context. Experiencing and using practical skills helps
develop the ability to use information in a variety of ways, thus enabling students to
apply their knowledge and understanding more readily.
• The integration of practical work into the teaching programme quite simply brings
the theory to life. Teachers often hear comments from students such as “I’m glad
we did that practical because I can see what the book means now.” and “It’s much
better doing it than talking about it.”.

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• Chemistry, physics and biology are by their very nature, practical subjects – both
historically and in the modern world. The majority of students who enter careers in
science need to employ at least basic practical skills at some time in their career.
For all students, whether they regard themselves as scientists or non-scientists,
the skills that they develop by doing practical work, hand-eye coordination skills,
communication, numeracy and problem solving skills, will prove to be useful
transferable skills throughout their future life.
• A practical course develops many cross-curricular skills including literacy,
numeracy, ICT and communication skills. It develops the ability to work both in
groups and independently and with confidence. It enhances critical thinking skills
and it requires students to make judgements and decisions based on evidence,
some of which may well be incomplete or flawed. It helps to make students more
self-reliant and less dependent on information provided by the teacher.
• The skills developed are of continued use in a changing scientific world. While
technological advances have changed the nature of practical procedures, the
investigative nature of practical science is unchanged. The processes of
observation, hypothesis formation, testing, analysis of results and drawing
conclusions will always be the processes of investigative science. The ability to
keep an open mind in the interpretation of data and develop an appreciation of
scientific integrity is of great value both in science and non-science careers.
• Practical work is not always easy and persistence is required for skills and
confidence to grow. Students often relish this challenge and develop a certain
pride in a job well done.
• The more experience students have of a variety of practical skills, the better
equipped they will be to perform well in the practical exams, both in terms of skills
and confidence. While it could be argued that the required skills could be
developed for papers 31 and 32 simply by practising past-papers, the all-round
confidence in practical ability will be greatly enhanced by a wider experience.
Similarly for paper 5, while it might be argued that planning, analysis and
evaluation could be taught theoretically, without hands-on experience of
manipulating their own data, putting their plans into action and evaluating their own
procedures and results, students will find this section difficult and will be at a
distinct disadvantage in the examination. Those students who can draw on
personal experience, and so are able to picture themselves performing the
procedure they are describing, or recall analysing their own results from a similar
experiment are much more likely to perform well than those with limited practical
skills.

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What are the practical skills required by this course?
This course addresses seven practical skills that contribute to the overall
understanding of scientific methodology. In a scientific investigation these would be
applied in the following sequence.
1 Planning the experiment
2 Setting up / manipulating apparatus
3 Making measurements and observations
4 Recording and presenting observations and data
5 Analysing data and drawing conclusions
6 Evaluating procedures
7 Evaluating conclusions
The syllabus shows how these seven skills are assessed and the structure is common
to all three sciences. The emphasis of the AS syllabus is on developing an
understanding and practice of scientific procedures, the collection of data, analysis and
drawing conclusions. It also starts to develop critical evaluation of procedures by
suggesting improvements to experimental procedures. In general students find the
performance of practical procedures and the collection of data more accessible than
analysis, whilst evaluation is least readily accessed. To enable access to these more
demanding skills, students need to understand why an experimental procedure is
carried out in a particular way so that they can recognise sources of error or limitations
which could affect the reliability of their results. Students will not be able to evaluate
until they can critically review a practical procedure.
The A2 syllabus builds upon the skills developed in AS and its emphasis is on the
higher level skills of planning, analysis and evaluating. In order to plan effectively,
students need to be able to evaluate procedures and critically assess results. This is
best achieved by the performance of practical exercises starting in AS with relatively
straightforward and familiar contexts and developed in A2 by the use of more complex
procedures and less familiar contexts. Data analysis again develops from AS into more
complex treatments so that students need to be given opportunities to gather suitable
data and perform the appropriate manipulations. The evaluation of conclusions and
assessing procedures are very high order skills. Students who have not had sufficient
opportunity to plan and trial their own investigations will find these skills difficult.
Students are not expected to be able to plan perfectly, but to recognise weaknesses
and make reasonable suggestions for improvement. The best learning tool to develop
these skills is to devise a plan, carry out the investigation and then assess how well the
planned procedure worked. The syllabus gives detailed guidance on the expected skills
and learning outcomes.
In summary, as the syllabus clearly shows, skills 2-6 listed above will be assessed at
AS level in papers 31 and 32. Skills 1 and 7 will only be assessed at A level in paper 5,
which will also take skills 5 and 6 to a higher level.
The above list shows the seven skills in the order in which they would be used in an
extended investigation. It is not suggested, nor would it be wise, to teach these skills in
this order. Students who are new to practical work will initially lack the basic
manipulative skills, and the confidence to use them. It would seem sensible, therefore,
to start practical training with skill 2, initially with very simple tasks and paying attention
to the establishment of safe working practices.
Once a measure of confidence in their manual dexterity has been established, AS
students can move on to exercises that require skills 3 and 4 to be included. Extensive

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experience in carrying out practical procedures allows students to gain awareness of
appropriate quantities and become more organised in time management and the
recording of data as it is collected.
It is likely that skill 6, Evaluating Procedures, will be the most difficult to learn at AS
level. Critical self-analysis does not come easily to many people. ‘My experiment
worked well’ is a frequent and inappropriate response. If students are to master this
skill, they need to develop an appreciation of reliability and accuracy inherent in the
equipment and procedure they are using. Only then will they be able to identify
anomalous results, or results which fall outside of the ‘range of uncertainty’ intrinsic in
the choice of apparatus used and so are considered to be inaccurate. Exercises with
less reliable/accurate outcomes can be used to provide more scope for the evaluation
of procedural, technique or apparatus errors.
Planning is arguably the most demanding of the seven skills. For it to be effective,
students need to be very well grounded in skills 2-6, so that they can anticipate the
different stages involved in the task, and can provide the level of detail required. It is for
this reason that planning skills are not assessed at AS level but form part of the A2
assessment in Paper 5. Unless students use apparatus they do not develop an
understanding of how it works and the sort of measurements that can be made using
particular sorts of apparatus. Candidates cannot be taught to plan experiments
effectively unless, on a number of occasions, they are required:
• to plan an experiment;
• to perform the experiment according to their plan;
• to evaluate what they have done.
The evaluation of conclusions, skill 7, is done by comparison of the outcome of an
exercise with the predicted outcome, and so is also an A2 skill. It should be taught and
practised as part of the planning exercises.

Summary of each of the seven skills

Full details of the requirements for each of these skills may be found in the Practical
Assessment section of the syllabus. What follows below is a brief summary of the
skills involved.
1 Planning
• Defining the problem
Students should be able to use information provided about the aims of the
investigation, or experiment, to identify the key variables. They should use
their knowledge and understanding of the topic under consideration to make a
quantitative, testable, prediction of the likely outcome of the experiment.
• Methods
The proposed experimental procedure should be workable. It should, given
that the apparatus is assembled appropriately, allow data to be collected
without undue difficulty. There should be a description, including diagrams, of
how the experiment should be performed and how the key variables are to be
controlled. Equipment, of a level of precision appropriate for the
measurements to be made, and quantities to be used should be specified. The
use of control experiments should be considered.
• Risk assessment
Candidates should be able to carry out a simple risk assessment of their plan,
identifying areas of risk and suggesting suitable safety precautions to be
taken.

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 • Planning for analysis, conclusions and evaluation
Students should be able to describe the main steps by which their results
would be analysed in order that that valid conclusions might be drawn. This
may well include the generation of a results table and the proposal of
graphical methods to analyse data. Also, they should propose a scheme for
the interpretation and evaluation of the results themselves, and of the
experimental procedure employed in obtaining those results. There should be
an indication of how the outcomes of the experiment would be compared with
the original hypothesis.
2 Setting up / manipulating apparatus
It is important that students are allowed sufficient time and opportunity to develop
their manipulative skills to the point where they are confident in their approach to
experimental science. They must be able to follow instructions, whether given
verbally, in writing or diagrammatically, and so be able to set up and use the
apparatus for experiments correctly.
3 Making measurements and observations
• Measuring/observing
Whilst successfully manipulating the experimental apparatus, it is crucial that
students are able to make measurements with accuracy and/or to make
observations with clarity and discrimination. Accurate readings of meters or
burettes and precise descriptions of colour changes and precipitates will make
it much easier for students to students to draw valid conclusions, as well as
scoring more highly in the test.
• Deciding on what measurements/observations to make
Time management is important, and so students should be able to make
simple decisions on the number and the range of tests, measurements and
observations that can be made in the time available. For example, if the
results of the first two titrations are in good agreement, there is no need to
carry out a third.
Students need to be able to make informed decisions regarding the
appropriate distribution of measurements within the selected range, which
may not always be uniform, and the timing of measurements made within the
experimental cycle. They should also be able to identify when repeated
measurements or observations are appropriate.
The strategies required for identifying and dealing with results which appear
anomalous should be practised.
4 Recording and presenting observations and data
An essential, but frequently undervalued, aspect of any experimental procedure is
the communicating of the results of the procedure to others in a manner that is
clear, complete and unambiguous. It is vital that students are well practised in this
area.
• The contents of the results table
The layout and contents of a results table, whether it is for recording numerical
data or observations, should be decided before the experiment is performed.
‘Making it up as you go along’ often results in tables that are difficult to follow
and don’t make the best use of space. Space should be allocated within the
table for any manipulation of the data that will be required.

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 • The column headings in a results table
The heading of each column must be clear and unambiguous. In columns
which are to contain numerical data, the heading must include both the
quantity being measured and the units in which the measurement is made.
The manner in which this information is given should conform to ‘accepted
practice’.
• The level of precision of recorded data
It is important that all data in a given column is recorded to the same level of
precision, and that this level of precision is appropriate for the measuring
instrument being used.
• Display of calculations and reasoning
Where calculations are done as part of the analysis, all steps of the
calculations must be displayed so that thought processes involved in reaching
the conclusion are clear to a reader. Similarly, where conclusions are drawn
from observational data, the key steps in reaching the conclusions should be
reported and should be clear, sequential and easy to follow.
• Significant figures
Students should be aware that the number of significant figures to which the
answer is expressed shows the precision of a measured quantity. Therefore,
great care should be taken with regard to the number of significant figures
quoted in a calculated value. The general rule is to use the same number of
significant figures as (or at most one more than) that of the least precisely
measured quantity.
• Data layout
Students should be able to make simple decisions concerning how best to
present the data they have obtained, whether this is in the form of tabulated
data or as a graph. When plotting graphs they should be able to follow best
practice guidelines for choosing suitable axis scales, plotting points and
drawing curves or lines of best fit. In drawing tables they should be able to
construct a table to give adequate space for recording data or observations.
5 Analysing data and drawing conclusions
This skill requires students to apply their understanding of underlying theory to an
experimental situation. It is a higher-level skill and so makes a greater demand on
a student’s basic understanding of the biology involved. Even when that
understanding is present, however, many students still struggle. The presentation
of a clear, lucid, watertight argument does not come naturally to most people and
so much practice in this area is recommended.
• Interpretation of data or observations
Once data has been presented in the best form for analysis of the results of
the experiment, the student should be able to describe and summarise any
patterns or trends shown and the key points of a set of observations. Further
values such as the gradient of a graph may be calculated or an unknown
value found, for example from the intercept of a graph.
• Errors
Students should be used to looking at an experiment, assessing the relative
importance of errors and where appropriate, expressing these numerically.
Students should be aware of two kinds of error.
i The ‘error’ that is intrinsic in the use of a particular piece of
equipment. Although we refer to this as an equipment error, we really

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 mean that there is a ‘range of uncertainty’ associated with
measurements made with that piece of equipment. This uncertainty
will be present no matter how skilled the operator might be.
ii Experimental error, which is a direct consequence of the level of
competence of the operator or of the effectiveness of the
experimental procedure.
• Conclusions
Students should learn to use evidence to support a given hypothesis, to draw
conclusions from the interpretation of observations, data or calculated values
and to make scientific explanations of their data, observations and
conclusions. Whatever conclusions are drawn, they must be based firmly on
the evidence obtained from the experiment. At the highest level, students
should be able to make further predictions and ask appropriate questions
based on their conclusions.
6 Evaluating procedures
Arguably, this is one of the most important, and probably one of the most difficult
skills for a student to develop. In order for the evaluation to be effective, students
must have a clear understanding of the aims and objectives of the exercise,
otherwise they will not be able to judge the effectiveness of the procedures used.
They must be able to evaluate whether the errors in the data obtained exceed
those expected due to the equipment used. If this is the case, they then need to
identify those parts of the procedure which have generated these excess errors,
and suggest realistic changes to the procedure which will result in a more accurate
outcome. Students should also be able to suggest modifications to a procedure to
answer a new question.
The evaluation procedure may include:
i the identification of anomalous values, deducing possible causes of these
anomalies and suggesting appropriate means of avoiding them,
ii an assessment of the adequacy of the range of data obtained,
iii an assessment of the effectiveness of the measures taken to control
variables,
iv taking an informed judgement on the confidence with which conclusions
may be drawn.
7 Evaluating conclusions
This is also a higher-level skill, which will demand of the student a thorough
understanding of the basic theory that underpins the science involved.
The conclusions drawn from a set of data may be judged on the basis of the
strength or weakness of any support for or against the original hypothesis.
Students should be able to use the detailed scientific knowledge and
understanding they have gained in theory classes in order to make judgements
about the reliability of the investigation and the validity of the conclusions they
have drawn.
Without practice in this area, students are likely to struggle. In order to increase the
confidence in drawing conclusions, it is recommended that practical exercises, set
within familiar contexts, be used to allow students the opportunity to draw
conclusions, make evaluations of procedure and assess the validity of their
conclusions.
In the examination, students may be required to demonstrate their scientific
knowledge and understanding by using it to justify their conclusions.

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Ways of doing practical work
Science teachers should expect to use practical experiences as a way to enhancing
learning. Practical activities should form the basis on which to build knowledge and
understanding. They should be integrated with the related theory, offering opportunities
for concrete, hands-on learning rather than as stand-alone experiences. In planning a
scheme of work it is important to consider a mosaic of approaches that include those
that allow students to participate in their own learning.
• Some practical activities should follow the well established structure that
includes a detailed protocol to follow. Such well-structured learning
opportunities have a vital role to play in introducing new techniques,
particularly in rapidly developing fields such as biotechnology. In these new
areas of science, teachers will often find themselves leading practical work
that they have not had the chance experience themselves as students.
• Other practical activities should offer the students the opportunity to devise
their own methods or to apply to solving a problem the methods that they have
been taught. The excitement generated by exposure to “new” and unfamiliar
techniques provides a stimulus to engage a student’s interest and challenge
their thinking.
Practical activities may be used as a tool to introduce new concepts – for example,
introducing catalysis by experimentation, followed up by theoretical consideration of the
reasons for the unexpected results obtained. On other occasions, practical work can
be used to support and enhance the required knowledge and understanding – for
example in building upon a theoretical consideration of the limiting factors of
photosynthesis with a series of practicals investigating the effect of light intensity and
hydrogen carbonate concentration on photosynthesis in water weed. In all cases,
learning will be enhanced most effectively by practical work that encourages students
to be involved, to think, to apply and use their knowledge, understanding and skills.
Practical work does not always have to be laboratory based. In classrooms, the use of
models, role play and paper cut-outs to simulate processes can be equally valuable.
Field studies also contribute greatly to a students’ appreciation of Biology and their
motivation and enjoyment of the subject. No amount of reading or viewing videos can
substitute for being exposed to an environment and the organisms living there. Even a
carefully managed environment like a school lawn represents a challenge to recognise
the species and to understand how they can survive.
There are a variety of strategies by which practical work can be integrated into a
scheme of work. Teachers should use a variety of methods, enhancing a variety of
subject specific skills and simultaneously developing a variety of transferable skills that
will be useful throughout their future professional lives. Some of the ways of delivering
practical work also enable the teacher to interact on a one-to-one basis with individual
students. This allows a teacher to offer support at a more personal level and develop a
greater awareness of an individual students needs.
Your choice of the specific strategy to use will depend on such issues as class size,
laboratory availability, the availability of apparatus, the level of competence of your
students, availability and expertise of technical support, the time available, your
intended learning outcomes for the activity and safety considerations. The following are
some possible strategies for delivery of practical work.

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• Teacher demonstrations
These require less time than a full class practical, but give little opportunity for
students to develop manipulative skills or gain familiarity with equipment.
Careful planning can give opportunity for limited student participation. Teacher
demonstrations are a valuable way of showing an unfamiliar procedure at the
start of a practical session, during which students go on to use the method.
Considerations in choosing to do a demonstration might include:
i Safety – some exercises carry too high a risk factor to be performed in
groups.
ii Apparatus – complicated procedures or those using limited resources
iii Time – demonstrations usually take less time
iv Outcome – some results are difficult to achieve and may be beyond the
skill level of most of the students. A failed experiment may be seen as a
waste of time.
v Students’ attention – a danger is that the attention of some students will
drift.
vi Manipulative experience – the teacher gets experience, the students’
don’t.
There are many good reasons for the teacher performing a demonstration but
do be aware that most students have a strong preference for hands-on
experimentation. So, where possible, do let them do it!
• Group work
Whole class practical sessions. These have an advantage in terms of
management as all the students are doing the same thing. Students may be
working individually, in pairs or in small groups. Integrating this type of
practical is straightforward as lessons beforehand can be used to introduce
the context and following lessons can be used to draw any conclusions are
develop evaluation. Where specialised equipment or expensive materials are
in short supply this approach may not be feasible.
Small group work. This can provide a means of utilising limited resources or
managing investigations that test a range of variables and collect a lot of
measurements. Although the same procedure may be performed, each
student group collects only one or a few sets of data which are then pooled.
For example, if five concentrations of the independent variable are being
tested, each of which need to be measured at two minute intervals for thirty
minutes, then a group of five students can each test one concentration. In
biology, field studies also lend themselves to group activities as a lot of data
has to be collected in a short period of time. The individual student has the
opportunity to develop their subject-specific skills. Part of the role of the
teacher is to monitor and maintain safety and also to enable and persuade
reluctant learners to take part. Group work aids personal development as
students must interact and work co-operatively.
Considerations might include:
i Learning – successful hands-on work will reinforce understanding; also,
students will learn from each other.
ii Confidence – this will grow with experience

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 iii Awareness/insight – should grow with experience
iv Team building – a most desirable outcome.
v Setting out – all students doing the same thing is easier for the
technicians
vi Confusion – incomplete, ambiguous or confusing instruction by the
teacher will waste time while the instructions are clarified but may also
compromise safety and restrict learning.
vii Opting out – some students will leave it for others to do and so learn very
little.
viii Safety – this could be a serious issue and constant vigilance is essential.
ix DIY – the urge to adapt their experiments, to ‘see what would happen if’,
must be strictly dealt with.
x Discipline – practical time must not be allowed to become ‘play time’.
Working in groups, whether as part of a whole-class situation or where groups
are working on parts of a whole, is probably the preferred option for many
students. At A level, it is highly desirable to include opportunities for students
to work on their own, developing their own skills and independence. In Papers
31 and 32, a student’s practical skills will be assessed on an individual basis,
so an individual’s experience, competence and confidence are of considerable
importance.
• Circus of experiments
A circus comprises of a number of different exercises that run alongside each
other. Individual or groups of students work on the different exercises and, as
each exercise is completed, move on to the next one. These are a means by
which limited resources can be used effectively.
There are two basic approaches. Most commonly, during a lesson a number
of short activities are targeted at a specific skill. Alternatively, over a series of
lessons, a number of longer practical activities are used, addressing a variety
of skills. The circus arrangement may be more difficult to manage as the
students are not all doing the same activity. This puts more pressure on the
teacher as they have to cope with advising and answering questions from a
variety of investigations. With circuses spread over a number of sessions,
careful planning is needed to enable the teacher to engage each group of
students, to maintain a safe environment. In these situations it is useful to
have at least two of the circus activities that involve no hands-on practical
work - using data response based simulations or other activities. In this way
the teacher can interact with groups that need a verbal introduction or short
demonstration and can monitor their activities more effectively.
i Apparatus – if the amount of apparatus used in an exercise is limited,
students are able to use it in rota.
ii Awareness – students by observing their peers will become more aware
of the pitfalls of the exercise and so will learn from the experience of
others.
iii Safety – different exercises may well carry different safety risks, all of
which would need to be covered.
iv Setting out –students doing different exercises will make it more difficult
for the technicians

11
 v Opting out – some students ay be tempted to ‘borrow’ the results of
earlier groups.
• Within theory lessons
This option should be considered whenever it is viable. It is likely that the
practical work would be by demonstration, as this would take less time. Given
the power of visual images, the inclusion of a short practical to illustrate a
theoretical point will reinforce that point and so aid the learning process. It is
critical, however, that the practical works correctly, otherwise the flow of the
lesson is disrupted and confidence in the theory may be undermined. The
exercise should therefore be practiced beforehand.
• Project work
Projects are a means by which a student’s interest in a particular topic, which
is not always directly on the syllabus, can be used to develop investigative
skills. It can also be used to access parts of the syllabus that have little
laboratory based investigation. For example, in gene technology students
might use internet based research to find examples of genetic modification
and present a poster display showing the implications. This sort of
investigative work can be individual, or a group activity. Once the project is
underway, much of the work can be student-based, outside the class room.
Care is needed in selecting the topics and setting a time scale, so that the
relevance is maintained to the syllabus context. The work can be directed at
the production of posters, presentations to give to the group or reports from
the group or individual.
Extra-curricular clubs
The role that these can play is in stimulating scientific enquiry methods. There
are a number of ways of using clubs. One way is to hold the club session
during the teaching day so that all students can attend. In effect this becomes
additional lesson time in which students can practice investigative skills,
including laboratory work. Such lab work involves materials that have a cost,
which must be planned for beforehand. If however the club is held outside the
teaching day it may be voluntary. Syllabus specific activities should be limited
and the most made of the opportunities for exciting work unrelated to
syllabuses. After school clubs could be vehicle for project work that is related
to science and of social or economic importance, for example, endangered
species or local mineral resources. Students who do attend the club could be
used as a teacher resource by bringing back their finding to a class room
session.
Keeping records
Students often find it a problem to integrate the practical work to the theory. This is
particularly true when a series of experiments or a long term investigation or project is
undertaken. Some potential issues include:
• Some students use odd scraps of paper in the laboratory, which are lost or
become illegible as chemicals are spilled on them. One important criterion is
that students are trained to record results immediately and accurately.
• Practical procedures may be provided, or students write their own notes from
a teacher demonstration. These may be lost, so students end up with results
but no procedure or context.
• When results take a period of time to collect, analysis becomes isolated from
the context of the investigation and may not be completed.

12
The key to minimising these issues is to train students into good work practices. This
is particularly important in colleges where students join at the start of their A levels from
a variety of feeder schools. It is also vital for students with specific learning difficulties
that affect their ability to organise their work such as dyslexia and Asperger’s
syndrome.
Students may be encouraged to integrate the practical in the same file as the theory.
Alternatively, students may be encouraged to keep an entirely separate practical book
or file. Loose leaf files make it easy to add to the file, but may make it easier to lose
items. Exercise books can be used but students should be encouraged to glue
provided protocols and their laboratory records into the book so that they are not lost.
Depending on how they learn, individuals may vary in their preferred method.
Whichever option is chosen, students need to be encouraged to relate their
investigations to the appropriate theory and to regard it as something that needs to be
thoroughly assimilated.
• Integrating the materials generated by practical work with the notes from
learning of theory can be achieved by interspersing the records of
investigations with the relevant section of theory. This may still require cross-
referencing where several learning outcomes and assessment objectives are
targeted by work.
• Keeping a separate practical book enables records of all the practical
investigations to be kept in one place. Students need training to manage
practical files effectively, particularly in keeping the contexts and cross
referencing to the theory. If care is not taken to develop and maintain these
skills, students may perceive practical as something different from theory.
• An intermediate between these two extremes is having a separate section for
practical investigations with each syllabus section in each student’s file and
cross-referenced to the relevant theory.
How is a practical activity organised?
Preparing for practical work needs thought and organisation. The practical work may
be an activity that forms part of a lesson, it may comprise an entire lesson, or it may be
an investigation designed to last for several lessons, but in every case, thorough
preparation is a key prerequisite to success.
Practical and investigative work should be integrated into the programme of study. The
scheme of work should identify appropriate practical investigative experiences for use
at the most suitable time. In designing the scheme of work,
• the resource implications should be considered in terms of equipment and
materials in stock,
• thought should be given to the seasonal availability of materials such as
organisms, and the sometimes short shelf-life of thermo-sensitive substances
such as enzymes or hygroscopic substances such as some salts,
• the time taken from order to delivery, potential for damage during despatch
and cost of materials to be obtained from local, national or international
suppliers should be considered,
• careful scheduling may be needed in Centres with a large number of students.
It may be possible to permit several groups to do the work simultaneously or in
quick succession, or it may be essential to re-order the scheme of work for
different groups so that scarce resources can be used effectively,

13
 • note must be taken of national or local health and safety regulations relating to
chemicals, electricity, growing microorganisms etc. There may also be
regulations controlling use of controversial materials such as genetically
modified organisms.
Once the scheme of work has been established, the next stage is to consider each
practical activity or investigation. In an ideal course, each of the following stages would
be gone through in developing each practical exercise in a course. This is not always
realistically possible the first time through a course, which is one reason for the
existence of this booklet. It is better to get going and to get some practical work done
with students than to hold out for perfection before attempting anything. Obviously, all
practical work should be subject to careful and rigorous risk assessment no matter how
provisional the rest of the supporting thinking and documentation.
• Decide on the aims of the work – the broad educational goals, in terms of the
broad skill areas involved (e.g. planning) and the key topic areas (e.g. animal
transport systems or unfamiliar material).
• Consider the investigative skills being developed. Reference should be made
to the syllabus, which in the practical skills section, includes learning
outcomes relating to practical skill. For instance, if the practical work intended
is to be a planning exercise, which of the specific skills identified in the
learning outcomes will be developed?
• With reference to the topics included, decide on the intended learning
outcomes of the practical activity or investigation, again referring to the
syllabus. For instance, which of the transport learning outcomes will be
achieved? In a few cases during the course, the material on which the
practical is to be based may be unfamiliar, in which case there may be no
topic-related intended learning outcomes. Thus, A2 contexts may be used for
AS practicals, and topic areas not on the 9700 syllabus at all may be used for
AS or A2 practicals.
• In addition, it may be useful to assess any other context of the practical work
investigation. For instance, is it intended as part of the introduction of a
concept, or to support a theory, or to demonstrate a process?
• Produce a provisional lesson plan, allocating approximate times to
introduction, student activities and summarising.
• Produce and trial a student work sheet. Published procedures or those
produced by other teachers can be used. Alternatively produce your own. As a
rule schedules produced by others need modifying to suit individual groups of
students or the equipment available. It helpful to ask students or another
teacher to read work sheets before they are finalised as they can identify
instructions that are ambiguous or use inaccessible terminology.
• Refine the lesson plan in relation to the number of students for which the
investigation is intended (whole class or a small group), the available
equipment (does some have to be shared?) and materials. There are
examples of lesson plans and student work sheets in appendix 2.
• Carry out a detailed and careful risk assessment (see below) before any
preparatory practical work is done, and certainly well before students do any
of the practical work. You should consider
o the likelihood that any foreseeable accident might occur – for example,
pupils putting glass tube through bungs are quite likely to break the tube
and push it though their hand

14
 o the potential severity of the consequences of any such accident – for
example dropping onto a desk a plastic dropper bottle of 0.01 mol dm-3
hydrochloric acid will cause much less severe eye injuries than the same
accident with a glass bottle containing 5.0 mol dm-3 hydrochloric acid.
o the means that can be taken to reduce the severity of the effect of any
accident – for example, the teacher or technician preparing bungs with
glass tubes before the lesson, or using eye protection such as safety
spectacles during all practical work.
• Make an equipment and materials list. This may need to be in sections;
o materials and apparatus per student or per group (chemicals and
glassware)
o shared equipment per laboratory (water baths, microscopes, pH meters)
o any chemicals should include concentrations and quantities needed
o any equipment should include number required
o any hazard associated with specific chemicals or equipment should also
be noted and cross referenced to the risk assessment. Sources of
information about safety may be listed in the syllabus (and are reproduced
below).
o The location of storage areas for equipment and chemicals may be cross
referenced to this equipment and materials list.
• Set up and maintain a filing system where master copies of the work sheets,
lesson plans and equipment lists can be stored. It is helpful to have these
organised, or at least indexed, by both their syllabus context and skills
developed.
• Once an investigation has been used by a group of students it should be
evaluated in relation to intended outcomes and the lesson plan. It is important
to obtain feedback from the students about their perception of the work. For
example,
o was the time allocation appropriate,
o were the outcomes as expected,
o did the students enjoy the work,
o did the students understand the instructions,
o was the point of the work clear to the students?
If necessary the work sheet and lesson plan should be revised.
Risk assessment
All practical work should be carried out in accordance with the health and safety
legislation of the country in which it is done. No activities should be attempted if they
conflict with such legislation.
Hands-on practical work can be carried out safely in schools. If it is to be safe, then the
hazards need to be identified and any risks from them reduced to insignificant levels by
the adoption of suitable control measures. These risk assessments should be done for
all the activities involved in running practical science classes including storage of
materials, preparatory work by the teacher and by any technical support staff and the
practical activities that are carried on in the classroom, whether demonstrations by the
teacher or practical activities for the students. Such risk assessments should be
carried out in accordance with the health and safety legislation of the country in which
they are done.
Risk assessment involves answering two basic questions:
1 how likely is it that something will go wrong? For example, pupils using a
double sided razor blade to cut up carrots are quite likely to cut themselves.

15
2 how serious would it be if it did go wrong? For example the consequences of
a spark from an experiment landing in an open bottle of magnesium powder are
likely to be serious, including spraying burning magnesium all over the laboratory,
burning many pupils and setting the laboratory ceiling on fire (based on a real
accident).
With the answers to these questions it is now possible to plan the practical activity to
minimise the risk of an accident and to minimise how severe any accident might be. In
our examples, this might include cutting up the carrot before giving to young pupils, or
providing older pupils with an appropriate sharp knife, it might include bringing in to the
laboratory only the amount of magnesium powder required for the activity.
How likely it is that something will go wrong depends on who is doing it and what sort
of training and experience they have had. You would obviously not ask 11 year old
students to heat concentrated sulphuric acid with sodium bromide, or to transfer
Bacillus subtilis cultures from one Petri dish to another, because their inexperience and
lack of practical skills makes a serious accident all too likely. By the time they reach
post-16 they should have acquired the skills and maturity to carry such activities out
safely.
Decisions need to be made as to whether an activity should be a teacher
demonstration only, or could be done by students of various ages. This means that
some experiments should normally only be done as a teacher demonstration or by
older students. Perhaps with well-motivated and able students it might be done earlier,
but any deviation from the model risk assessment needs discussion and a written
justification beforehand.
There are some activities that are intrinsically dangerous, and, if included in the
suggested activities, should always be changed to more safe modes of practice, for
example, there are no circumstances under which mouth pipetting is acceptable –
pipette fillers of some sort should always be used.
Teachers tend to think of eye protection as the main control measure to prevent injury.
In fact, personal protective equipment, such as goggles or safety spectacles, is meant
to protect from the unexpected. If you expect a problem, more stringent controls are
needed. A range of control measures may be adopted, the following being the most
common. Use:
• a less hazardous (substitute) chemical;
• as small a quantity as possible;
• as low a concentration as possible;
• a fume cupboard; and
• safety screens (more than one is usually needed, to protect both teacher and
students).
The importance of lower concentrations is not always appreciated, but the following
examples, showing the hazard classification of a range of common solutions, should
make the point.
ammonia (aqueous) irritant if ≥ 3 mol dm-3 corrosive if ≥ 6 mol dm-3
sodium hydroxide irritant if ≥ 0.05 mol dm-3 corrosive if ≥ 0.5 mol dm-3
hydrochloric acid irritant if ≥ 2 mol dm-3 corrosive if ≥ 6.5 mol dm-3
nitric acid irritant if ≥ 0.1 mol dm-3 corrosive if ≥ 0.5 mol dm-3
sulphuric acid irritant if ≥ 0.5 mol dm-3 corrosive if ≥ 1.5 mol dm-3
barium chloride harmful if ≥ 0.02 mol dm-3 toxic if ≥ 0.2 mol dm-3
(or if solid)

16
Reference to the above table will show, therefore, that if sodium hydroxide is in
common use, it should be more dilute than 0.5 mol dm-3. The use of more concentrated
solutions requires measures to be taken to reduce the potential risk.
Material Safety Data Sheets (MSDS)
Your risk analysis should consider the hazards associated with the materials you
propose to use. These risks are best assessed by reference to MSDS’s appropriate to
the chemical(s) in use. These are generally supplied by the chemical manufacturer and
supplied with the chemical. If this is not the case then there are many internet sites that
have this information freely available. These sheets also provide useful information on
the actions to take following an accident, including first aid measures, and should
therefore be considered essential for all practical experiments involving chemicals, as
part of the risk assessment process.
Hazard key
The following key applies.
C = Corrosive substance F = Flammable substance
H = Harmful or irritating substance O = Oxidising substance
T = Toxic substance N = Harmful to environment

Eye protection
Clearly students will need to wear eye protection. Undoubtedly, chemical splash
goggles give the best protection but students are often reluctant to wear goggles.
Safety spectacles give less protection, but may be adequate if nothing which is classed
as corrosive or toxic is in use.
Your risk assessment should not restrict itself simply to the materials, procedures and
equipment being used, but should have a wider remit, covering the time from when the
class enter the room until they leave it.
Practical science can be - and should be - fun. It must also be safe. The two are not
incompatible.
Safeguards in the School Laboratory, 10th edition, ASE, 1996
Topics in Safety, 2nd edition, ASE, 1988
Hazcards, CLEAPSS, 1998 (or 1995)
Laboratory Handbook, CLEAPSS , 1997
Safety in Science Education, DfEE, HMSO, 1996
Hazardous Chemicals Manual, SSERC2, 1997.

17
AS Skills
AS skills will form the foundation on which A2 skills will be developed. Students will
become competent in these skills through practical experience. They should be
expected, during the AS course, to carry out as much practical work as possible, since
this with develop both key practical skills and enhance their motivation as well as their
understanding of the theory part of the course. The specific investigations to which
references are made can be found in appendices 1 and 2. The syllabus clearly
describes the skills that are to be assessed, and should be used to ensure that
activities are appropriately targeted.

Teaching students to manipulate, measure and observe

As part of their AS studies students will be expected to develop skills in manipulating
and measuring using standard laboratory apparatus. These will form a basis on which
more advanced manipulative skills will be developed in A2. During their AS course it is
assumed that students will learn how to measure accurately and to manage their time
effectively, so that they are confident in their use of apparatus.
• A good starting point to practice these skills is with microscope work (e.g.
practicals 1 and 2). Students will be expected to be confident in the use of
microscopes and be able to make temporary mounts of a variety of
specimens. It is most important that they are capable of recording their
observation by being able to make clear, well proportion, labelled drawings of
what they observe. This is not an easy skill to acquire and time should be
spent to ensure that students develop their capability in this skill.
• Various investigations (such as practicals 5 and 6) will also allow students to
collect data and make observations. This will require that students are able to
follow a set of instructions and set up apparatus appropriately. They should
then be able to collect data using a wide variety of means.
• Students should be able to make informed decisions about the number of
times a reading should be taken and the range of readings that is required to
collect reliable and valid data. Students should also be able to replicate
readings or observations as necessary. Many of the practical investigations
here offer such opportunities, but particularly open-ended investigations such
as practical 16.

Teaching students to present data and record observations

Many students do not find this an easy skill to master. It is important that students can
record data so that it is capable of being understood by others. This requires skill in
deciding how to present the data and what should be recorded.
• Students need to be able to present numerical data in tabular form and to
decide on the structure of the table and what titles and units should be written
in the column headers. They should produce the table so that readings from
the investigation may be entered directly into the table as the readings are
taken. Space should be allocated in tables as necessary for calculated values
and deductions. Opportunities include practical 7 and 19.
• Students should ensure that all readings are taken with the same degree of
accuracy and precision.
• Students often assume that everyone understands how they achieved their
answers to questions or calculations without realising that this is not the case.

18
 This is particularly true when answering examination questions. Examiners
can only give credit for what they see and students may well receive credit for
a correct method even if they reach the wrong answer or conclusion. However
this requires that the students display their calculations and reasoning.
• Students should show the working in their calculations and the key steps
in their reasoning.
• Students should also use the correct number of significant figures for
calculated quantities.
Several of the practicals include such numerical work, for example practicals
2, 13, 14 and 20.
• Students should be able to choose a suitable method of presenting data
obtained from an investigation for example quantitative data as graphs (e.g.
practicals 6 and 7), qualitative data as tables (e.g. practicals 3 and 4) and
cellular and histological data as drawings (e.g. practicals 1, 2, 13 and 14).
• When producing graphs, students should be able to select which variables to
plot on the x and y axes. They should be able to plot with accuracy and follow
the Institute of Biology recommendations for drawing lines on graphs.
• More information concerning the presentation of data and observations is
provided in the syllabus.

Teaching students to analyse, draw conclusions and evaluate

These are the hardest skills that have to be mastered by students. Evaluation in
particular is found very difficult by most students as they are having to think in the
abstract rather than handle real apparatus and materials. It is most important that the
basics in these skills are mastered so that they can be further developed in the A2 part
of the course.
• Students need to be able to interpret data or observations by describing
patterns and trends shown by tables and graphs. In data such as highly
curved graphs, the key patterns should be described (e.g. practical 5) and in
data producing simple curves or straight lines, the trend of the data observed
and described (e.g. practical 6).
• Students should be able to determine unknown values by extrapolation and
interpolation of lines on graphs and be able to calculate the mean from
replicated observations.
• It is most important that students are able to explain the degree of confidence
they have in their conclusion and identify and explain possible sources of error
in the investigation.
• Students should be able to say whether the data obtained supports the
original hypothesis and use this to make further predictions.
• The ability to make simple evaluations should be practiced so that this skill
can be further developed in A2. This enables students to suggest
improvements to procedures so as to improve the reliability of the data
obtained and to extend investigations into new situations or solve related
problems. The more practice students have of this skill, the better. Ideally
every investigation could be evaluated using a simple check list until it
becomes an automatic response by a student.

19
 Appendix 1

Appendix 1 - Designing a practical course for AS

Outline List of Practical Experiments
Full details are provided for practicals 1 - 11
Number and title type of syllabus
practical reference
1 Size and scale obs A
2 Plant tissue observation obs A
3 The identification of biological chemicals present in solutions wet B
4 The metabolism of different carbohydrates by yeast wet B
5 Effect of pH on enzymes wet C
6 Effect of inhibitors on enzymes wet C
7 Using beetroot to investigate cell membrane permeability wet D
8 Root tip squash wet E
9 The extraction of DNA from onions wet F
10 Effect of wind-speed on transpiration rate using a potometer wet G
11 Investigating the role of carbon dioxide in living organisms wet H
12 Size and scale obs A
13 Plant tissue observation obs A
14 Animal tissue observation obs A
15 Food tests wet B
16 Effect of temperature on enzymes wet C
17 Effect of substrate concentration on enzymes wet C
18 Effect of enzyme concentration wet C
19 Osmosis wet D
20 Plasmolysis and water potential w&o D
21 Chromosome observation - prepared root tip squash obs E
22 Stomata obs G
23 Effect of temperature and light on transpiration rate wet G
24 Investigate transport of water using celery wet G
25 Investigate tidal and vital capacity wet H
26 Effect of nicotine on heart rate of daphnia wet H
27 Bacteria and viruses obs I
28 Blood cell smear w&o J
29 Food chains and food webs field K
30 Nutrient cycling wet K
The route which teachers take through the AS syllabus may vary. The practical
course follows the sequence of the syllabus, although some learning outcomes may
well be grouped differently. Teachers can adapt the investigations as they wish as
they wish. All of the investigations that are supplied in detail are intended to be
completed within a one hour time span.
CIE 9700/03 practical papers are not intended to be used for formative development
of practical skills. The questions set for the previous syllabus are written to different
assessment objectives, although some of the practical components could be used as
practice exercises. Teachers who choose to use past papers as a student’s only
learning experience are placing these students at a disadvantage.

20
 Appendix 1

Syllabus section A – Cell Structure

Practical 1 – Size and scale 1
Learning outcomes (a) use a stage micrometer and graticule and (f) calculate
linear magnification of drawings
Practical skills: Manipulation, measurement and calculation.

The practical uses a stage micrometer to calibrate an eyepiece graticule so that

specimen size can be determined. The candidates can also determine the scale of
drawings that are made of the specimens under observation. It is suggested that the
eyepiece lenses be fitted with the graticules prior to the investigation.

Full details of this practical are provided.

Practical 12 – Size and scale 2 The investigation described in practical 1 uses a TS

of Lamium stem, but the practical can and should be modified and form a template
for a series of practicals to include other examples from a wide range of plant and
animal tissues.
Practical 2 – Plant tissue observation
Learning outcomes (f) draw plan diagrams of tissues. The practical can be
extended to also cover (b) (e) and (g) distinguish between resolution and
magnification and compare and contrast structure of plant and animal and
prokaryotic and eukaryotic cells.
Practical skills: Observation and recording and interpreting observations.

The practical emphasises and practices microscopic and drawing skills so that
candidates are able to use a microscope and produce clear, well proportioned,
labelled drawings in both low and high power. They also learn the difference between
cellular and plan drawings and how best to represent the various tissues of a
specimen under observation.

Full details of this practical are provided.

Practical 13 – Plant tissue observations Practical 14 – Animal tissue

observations The investigation described in practical 2 should be modified and form
a template for a series of practicals at appropriate points through the course, to
include other examples from a wide range of plant and animal tissues.

Syllabus section B – Biological Molecules

Practical 15 – food tests – students may well have used food tests in their previous
biology courses, but they should be given an opportunity to use Benedict’s test for
reducing and non-reducing sugars, biuret test for protein, iodine in potassium iodide
solution for starch and the ethanol emulsion test for lipid. They should be given
prepared solutions / oils to try, as well as liquid food materials such as milk and food
materials that need grinding up and suspending such as bread.

21
 Appendix 1

Practical 3 – Identification of biological chemicals present in solutions

Learning outcomes (a) carry out tests for reducing and non-reducing sugars,
starch, lipids and proteins.
Practical skills: Manipulation, measurement, observation, recording of data,
interpreting and drawing conclusions.

The practical develops an understanding of the application of different biochemical

tests to the problem of identification of unknown substances. The practical
concentrates on developing skills of decision making, recording, drawing conclusions
and identifying alternative strategies.
Full details of this practical are provided.
Practical 4 – the metabolism of different carbohydrates by yeast
Learning outcomes (a), (b) and (d) form the context for this practical, making
use of unfamiliar methods and materials and thus (e) use the knowledge
gained to solve related problems in a new situation.
Practical skills: Manipulation, measurement and observation, presentation of
data and analysing and forming conclusions.

The practical investigates how efficient yeast is in metabolising different

carbohydrates by observing the time taken for methylene blue to be discoloured. The
ideas developed can include previous work done on cell membranes and the
different ways that substances are transported into and out of cells.

Syllabus section C – Enzymes

Practical 5 – Effects of pH on enzymes
Learning outcomes (c) follow the course of an enzyme catalysed reaction and
(d) investigate and explain the effect of pH on the rate of enzyme catalysed
reactions.
Practical skills: Manipulation, measurement and observation, presentation of
data and analysing and forming conclusions.

The practical investigates the effect of pH on enzymes and the effect on the
hydrogen and ionic bonds that determine the shape of the active site. Candidates
may determine the optimum pH and the point at which the enzyme has been
denatured.

Full details of this practical are provided.

Practical 6 – Effect of inhibitors on enzymes
Learning outcomes (c) follow the course of an enzyme catalysed reaction and
(e) explain the effects of a non-competitive inhibitor on the rate of enzyme
activity.
Practical skills: Manipulation, measurement and observation, presentation of
data and analysing and forming conclusions.

The practical investigates the effect of a non-competitive inhibitor, lead nitrate, on the
hydrolysis of starch.

Full details of this practical are provided.

22
 Appendix 1

Practical 16 – Effect of temperature on enzymes The effect of temperature on

enzyme catalysed reactions should be investigated. Candidates can carry out a
simple enzyme catalysed reaction at different temperatures using provided troughs /
plastic bowls at different temperatures as water baths. This works well using
protease such as trypsin or bacterial protease and exposed, developed black-and-
white photographic film, or using amylase and starch. The data collected can then
be displayed graphically to assist in drawing conclusions. Experiments of this type
lend themselves to be critically analysed, evaluated and improved.
Practical 17 – Effect of substrate concentration on enzymes Practical 18 effect of
enzyme concentration These experiments can be further modified to investigate the
enzyme concentration and the concentration of substrate on the rate of reaction.
This would offer the opportunity to use other enzyme/substrate systems such as
urea/urease (detect time taken for ammonia to be produced), yeast
catalase/hydrogen peroxide (height of foam or count bubbles from delivery tube or
collect gas over water and measure volume) or lipid emulsion/lipase (detect changes
in pH). In addition this will also practice the skills of identifying and controlling
variables. At this level, candidates should also be able to identify dependent and
independent variables.

Syllabus section D – Cell Membranes and Transport

Practical 7 – Using beetroot to investigate cell membrane permeability
Learning outcomes (b) outline the role of membranes within cells, (c) describe
the process of diffusion and (e) use the knowledge gained to solve related
problems in a new situation.
Practical skills: Manipulation, measurement and observation, presentation of
data and analysing and forming conclusions.

The practical investigates the effect of temperature on the permeability of cell

membranes. Candidates will need access to a colorimeter for this experiment
although it could be modified so that diluted solutions were compared against a
standard colour chart. Paint manufactures produce a very wide range of standard
colours for paints mixed ‘in store’ and a selected sample of these could be used for
‘standards’.

Full details of this practical are provided.

Practical 19 – Osmosis This can be investigated in a variety of ways such as
determining the water potential of potato tissue by placing samples of potato in
different concentrations of sucrose solution and noting the gain or loss in mass of the
tissue. The data can be displayed graphically and the concentration determined by
interpolation on the graph.
Practical 20 – Plasmolysis and water potential Plasmolysis can be investigated in
epidermal onion tissue. The water potential can be determined by finding incipient
plasmolysis in cells placed in different concentrations of solution. This extends the
work done in the previous practical on osmosis by including the skills of microscope
technique in the practical. Students may well need training in order to identify cells
with incipient plasmolysis.

23
 Appendix 1

Syllabus section E – Cell and nuclear division

Practical 8 – root tip squash
Learning outcomes (d) describe with the aid of diagrams, the behaviour of
chromosomes during the mitotic cell cycle.
Practical skills: Manipulation, observation, interpretation and recording
observations.

The practical involves preparing and staining cells from an active meristem in the
roots of plants. The cell tissue is then ‘squashed’ and observed using standard
microscopic techniques and different stages of mitotic cell division identified and
drawn. It is advised that due to shortness of time within a one hour lesson, that
candidates have previously done other microscope practicals so that they are familiar
with and can use a microscope with skill and precision.

Full details of this practical are provided.

Practical 21 – chromosome observation – prepared root tip squash Practical 8

can be further extended and modified so that candidates are provided with ready
made prepared cell tissue showing a full range of the different stages of cell division
in order that they can become skilful in identifying and drawing them. This is good
material for developing skills of measurement, observation and drawing as the
structures are intra-cellular and therefore small.

Syllabus section F – Genetic Control

Practical 9 – Extraction of DNA from onions
Learning outcomes (a) describe the structure and importance of DNA
Practical skills: Manipulation and making observations.

The practical involves the extraction of DNA from living material such as onion and
clearly and powerfully demonstrates to students the actual existence of DNA when it
has probably been experienced by the student as a theoretical substance found only
in textbooks.

Full details of this practical are provided.

Syllabus section G – Transport

Practical 10 – effect of wind-speed on transpiration rate.
Learning outcomes (c) describe how to investigate experimentally the factors
that effect transpiration rate.
Practical skills: Manipulation, data collection and analysis and drawing
conclusions.

The practical investigates transpiration in a leafy shoot using a potometer and the
effect that wind speed has on the rate of transpiration. Wind-speed is determined by
a fan that is set at different distances from the plant. Students come to realise that
this does not involve the inverse square law but this idea can be developed to show
how the inverse square law affects light and photosynthesis.

Full details of this practical are provided.

24
 Appendix 1

Practical 23 – Effect of light and temperature on transpiration rate. The

investigation in practical 10 can be modified so that others variables such as
temperature and light can be investigated to see how they affect transpiration rate.
Practical 22 – Stomata The structure of stomata can be observed by coating the
lower surface of a leaf with clear nail varnish, which is then pealed of and made into
a temporary slide for viewing through a microscope. This practical not only
investigates stomatal structure but also reinforces microscope skills learnt earlier in
the course. Leaves of different plants can be compared so that xerophytic
adaptations can be studied.
Practical 24 – Investigate transport of water using celery Transport of water
through plant tissue can investigated using celery and broad bean seedlings. A stick
of celery left to stand in a dilute solution coloured dye such as food colouring or ink
can be used to show the presence of vascular bundles in roots, stems and petioles.
Students can section the stem to show the distribution of the vascular bundles. The
veins in the leaves of the celery and been seedling will also become stained and thus
show what happens to the vascular bundles when they enter leaves. This enables
students to build up a whole picture of how the water is transported throughout the
plant.

Syllabus section H – Gas Exchange

Practical 25 – tidal and vital capacity Tidal and vital capacity can be investigated
using spirometer. Schools that do not possess a spirometer can do a simplified
version of this experiment by getting students to blow through rubber tubing placed in
a 1000 ml measuring cylinder or larger, full of water and inverted into a large bowl.
Measurements of expired air can be read of directly from the measuring cylinder.
Practical 26 – Effect of nicotine on heart rate of Daphnia The effect of nicotine
on the heart rate in daphnia can be investigated to show how different drugs can
affect metabolism.

To make the nicotine solution, remove tobacco from a pack of 20 cigarettes and mix
it with a beaker of water. Cover and allow to stand for at least 12 hours. Drain the
supernatant liquid and mix one part solution to eight parts water. Different
concentrations may be used to investigate the effect of concentration on heart beat.
Practical 11 – Investigating the role of carbon dioxide in living organisms
Learning outcomes (k) use the knowledge gained in this section in new
situations or to solve related problems
Practical skills: Manipulation, observation, interpretation and planning.
This practical investigates the carbon dioxide levels produced by living organisms
using bicarbonate indicator solution. The students then use this information to plan
an investigation to determine the unfamiliar interaction between respiration
(producing carbon dioxide at a constant rate irrespective of light intensity) and
photosynthesis (using carbon dioxide – dependent on light) in plants.

Full details of this practical are provided.

25
 Appendix 1

Syllabus Section I – Infectious disease

Practical 27 – Bacteria and viruses There are various ways of making practical
material on these accessible, but the most satisfactory will be to use electron
micrographs (downloaded from the web or in books) to make observations and
measurements.

Syllabus Section J – Immunity

Practical 28 – Blood cell smear – this must be done using prepared slides. This is
an important practical as the cells are small and not very easy to see, so they make
an excellent context for experienced students to finely tune their skills of using
apparatus, observation and drawing.

Syllabus Section K – Ecology

Practical 29 – food chains and food webs – students should have the opportunity
to go out into the school grounds or a suitable local environment in which are found
plants, animals that feed on the plants and predators (this might include trees, aphids
and carnivorous beetles, or herbaceous plants, snails and snail-eating birds).
Students should be enabled to identify, count (per unit area) and estimate biomass,
preparatory to making food webs and attempting to make a simple estimated pyramid
of biomass.
Practical 30 – nutrient cycling – students should set up small, sealed jars
containing a few grams of soil, pond water and a small amount of shredded paper.
These should be left in the light and observed for a few weeks. There will be a
succession of changes in the microorganisms growing in the jars, but eventually it will
settle down to a constant population of various photosynthetic and other species,
demonstrating that they are recycling the limited amount of nutrients present.

26
 Appendix 2

Appendix 2 – practicals for which full details are provided

Practical 1 - Investigation into size and scale of microscopic tissues

This practical focuses on microscope technique and using graticules and stage
micrometers to determine size and scale in biological cells and tissues.

Intended learning outcomes

By the end of this practical you should be able to:

• Use a microscope fitted with an eyepiece graticule and stage micrometer
• Calibrate the eyepiece graticule using the stage micrometer
• Use the calibrated graticule to determine the actual size of microscopic
specimens
• estimate the accuracy of a measurement
• Use the graticule to determine scales
• Understand the importance of repeating or validating set of results.
Safety Information

There are no particular hazards in this practical, however you must follow your
laboratory rules.

Background information
• The measurement of specimen size with a microscope, is made by using
an eyepiece graticule. This is a glass or plastic disc with 8 divisions etched
onto its surface, which is inserted into the eyepiece lens.
• The size of the eyepiece graticule remains constant, despite the fact that
the image viewed will change its size depending upon whether high- or
low-power objective lenses are used. For example a cell viewed with the
x40 objective will appear much larger than when viewed with the x10
objective. However because the graticule is in the eyepiece it will not
change its size. Therefore the value of each of the divisions in the
eyepiece graticule varies with the magnification of the objective lens.
• A stage micrometer is a very accurately etched glass or plastic ruler that is
placed on the microscope stage so that the eyepiece graticule scale is
superimposed on the stage micrometer scale. The scale is usually 1mm
divided into 100 separate divisions so that each division equals 10
micrometres (10µm).
• It is necessary to calibrate the eyepiece graticule with the stage
micrometer placed on the microscope stage for each objective lens used.

You will observe a TS of plant tissues through a microscope and use an eyepiece
graticule and a stage micrometer to determine the size of some of the structures.

• Read the information above.

• Ensure that you understand the principles of using an eyepiece graticule
and a stage micrometer before you continue with the investigation.

27
 Appendix 2

Method

Preparation
1. You have been provided with a compound light microscope with both low-
and high-power objective lenses and an eyepiece lens that has been fitted
with a graticule. You have also been provided with a stage micrometer.

eyepiece lens
fitted with graticule

objective lens

stage

2. You must now calibrate the eyepiece graticule.

Place the stage micrometer onto the microscope stage and focus using the
low-power objective lens so that the graticule scale becomes
superimposed over the stage micrometer scale.
points of coincidence on
stage micrometer scale

0 2 4 6 8
eyepiece graticule scale
3. Move the stage micrometer until the start or zero line of each scale is
coincident (lined up)
4. Look along the scale until another coincident point is found.
5. The relationship between the two scales can now be calculated
On the scale shown there are 17 divisions on the stage micrometer scale
that line up with 7 divisions on the graticule scale.
Thus 17 / 7 = 2.42857 units.
Each unit on the stage micrometer scale is 10 micrometres (10µm).
Therefore each division on the graticule scale is 24.2857 micrometres
rounded to 24.3 µm.
6. Use the procedure described above to determine the size of each division
on the eyepiece graticule using the low-power objective lens of your
microscope.
7. Repeat the procedure to determine the size of each division when using
the high-power objective lens.

28
 Appendix 2

Making observations
1. You are provided with a stained transverse section through part of a
dicotyledonous plant root.
2. Examine the specimen using the low-power of your microscope.
3. Make a large, plan drawing to show the distribution of tissues, labelling the
stele (vascular bundle).
4. Use the eyepiece graticule to measure the width of the vascular bundle at
its widest point in graticule units and then calculate the actual width of the
vascular bundle in millimetres and in micrometres.
5. Draw a straight line on your drawing across the vascular bundle to show
where you took your measurement. Write the dimension on your drawing
next to the line.
6. Make a high-power drawing to show a group of four xylem vessels from
inside the vascular bundle.
7. Use the eyepiece graticule to measure the width of the xylem vessel at its
widest point in graticule units and then calculate the actual width of the
vessel in micrometres, remembering to use the appropriate calibration of
the eyepiece graticule for the high-power objective lens.
8. Draw a straight line on your drawing across the xylem vessel to show
where you took your measurement. Write the dimension on your drawing
next to the line.
9. Look at your two measurements and check on their accuracy. The actual
size of the xylem vessel should be smaller than the size of the vascular
bundle even though it looked larger using the high power objective lens.
10. You are now going to determine the magnification of your drawing of the
xylem vessels. Use a ruler to measure the length of the line that you drew
across the xylem vessel.

Use your knowledge of the actual size of the vessel to calculate the magnification of
your drawing. Write your answer x at the bottom right hand corner of your
drawing.

Follow-up
• Compare your results with other members of the class and check for
consistency of readings.
• Did any member of the class have anomalous results? What are the
potential causes of such an anomalous result in this investigation?
• Write up your procedure including a discussion of the benefits of
comparing your results with other students.

29
 Appendix 2

Practical 1 - Lesson Plan

Investigation into size and scale of microscopic tissues.

Context

A practical investigation set in the context of 9700 syllabus – Assessment objective C

2 and 3 – Cell structure.

Key aims of the lesson

This practical is designed to develop the skills of using an eyepiece graticule and a
stage micrometer with a microscope to determine size and scale of microscopic
tissues.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Use a compound light microscope with an eyepiece graticule
• Use a compound microscope with a stage micrometer
• Calibrate an eyepiece graticule with a stage micrometer
• Use the calibrated graticule to determine the actual sizes of cells and tissues
using both low- and high-power objective lenses.

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – Student worksheet given out for students to read in

previous preparation for the practical lesson.
lesson

0-3 Introduction to the aims, intended outcomes and shape of the lesson
– teacher led oral presentation

3 - 10 Context – review the method for setting up and using a microscope

with an eyepiece graticule and stage micrometer and how to calibrate
the eyepiece graticule.

10 - 15 Introduction to method – Teacher briefly outlines method and

answers any student questions on procedure. Teacher emphasises
safety concerns regarding breaking slide by focussing down with the
objective lens.

30
 Appendix 2

15 - 40 Carrying out the practical – students carry out the practical work.

40 - 50 Obtain results – Students compare their results with other students

in the class

50 - 60 Drawing together the threads – Teacher led discussion on the skills

that have been developed as well as discussion on results obtained.

31
 Appendix 2

Practical 1 - Technical information

Investigation into size and scale of microscopic tissues.

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. 1 x prepared slide TS Ranunculus root
2. 1 x microscope with an eyepiece graticule fitted, suitable illumination and:
• high power objective lens e.g. x40 (equal to 4mm or 1/6”)
• low power objective lens e.g. x10 (equal to 16mm or 2/3”)
3. 1 x stage micrometer – a scale on a glass slide
4. suitable white paper, HB (medium-hard) pencil and rubber

Safety Precautions.
No specific hazards have been identified in this practical, however a risk assessment
should be carried out as a matter of course.

32
 Appendix 2

Practical 2 - Microscopic observation of cells and tissues

This practical focuses on microscope technique and making and recording

observations in the form of biological drawings.

Intended learning outcomes

By the end of this practical you should be able to:

• Use a microscope with skill and precision
• Show all the structures that can be seen in the defined part of a specimen
• Make clear, accurate, labelled, scale drawings of specimens

Safety Information.
There are no particular hazards in this practical, however you must follow your
laboratory rules.

33
 Appendix 2

Background information
• Drawings should be done with a sharp HB pencil making clear single lines.
Examiners do not give credit for sketchy lined drawings. A soft rubber can
be used to correct errors.
• Always draw what you see and not what you expect to see from memory
or textbook diagrams.
• Candidates often draw diagrams too small but rarely draw them too large.
Ensure that your drawing is large enough to show all the detail.
• All parts of the drawing should be kept in correct proportions. In poor
quality drawings, proportions changes as the drawing progresses.
• Biological drawings can be both high-power and low-power.
• Low-power drawings are usually plan drawings that do not contain cellular
detail but do show the distribution of various tissues. When a plan drawing
is requested, examiners may give credit for not drawing cellular detail.
• If more than one drawing of the same or different specimens or parts of a
specimen are made, examiners may ask that they are drawn to the same
scale (which means the same magnification). Credit is then awarded for
this skill.
• Look at the following two sets of drawings of a red and white blood cell,
made by different students and how marks would be allocated by an
examiner.

• Student A would be awarded 1 mark. Student B would be awarded 6 marks.

You will observe a TS of plant tissues through a microscope using both low and high
power and draw appropriate structures.

34
 Appendix 2

• Read the information above.

• Read your textbook and look carefully at any drawings that have been
made or biological material. Take care however; the quality of drawings in
some textbooks is not all that could be desired.
• Write down the key features that are found in good biological drawings.

Method

Preparation
1. You have been provided with a compound light microscope with both low-
and high-power objective lenses and a slide of a TS of a plant stem.

eyepiece lens

objective lens

stage

light source

2. Place the slide onto the stage of the microscope.

3. Adjust the light source so that you can see a bright light when looking
through the eyepiece lens.
4. With some microscopes it is possible to rack the objective lens so far down
that it will break the slide. In order to prevent this it is good microscope
technique to:
• set the objective lens on low power.
• not look through the eyepiece but to look at the side of the microscope
and carefully lower the objective lens until it is nearly, but not quite
touching the slide.
• now look through the eyepiece and gradually raise the objective lens
until the slide comes into focus.
5. You should now carefully move the slide around on the stage until you find
the area that you wish to observe.
6. To change to high power, do not re-focus, but change the objective lens
from low to high power. The slide should be almost in focus and only a fine
adjustment to the focus should be necessary.
7. Practice focussing the slide on both low and high power until you are
familiar with the technique.

35
 Appendix 2

Making observations
1. You are provided with a stained transverse section through part of a
dicotyledonous plant.
Examine the specimen using the low-power of your microscope.
Make a large, labelled, plan drawing to show the distribution of tissues.
2. Make a high power drawing to show a group of four cells from the region
nearest the centre of the specimen.

Follow-up
• State from which part of the plant the section was taken. Explain your
answer
• Exchange your drawings with another student and mark their drawings
using the following mark scheme.

Mark scheme

Plan drawing
Corner vascular bundles larger than other vascular bundles 9
No individual cells drawn 9
Four sided shape to plan 9
Both xylem and phloem correctly labelled 9
Parenchyma correctly labelled 9
Sclerenchyma on outer edge of vascular bundle labelled 9
Collenchyma in corners labelled 9

High power drawing

Good quality of drawing i.e. clear single lines 9
4 cells only drawn, similar in size and shape 9
between 5-8 sides to each cell 9
Air spaces shown between corners of cells 9
Thin cell walls shown either by a thin
single line or two lines close together 9
• Add up the marks out of 12 and return the drawings to the student.
• Write a list of all the reasons why you did not score full marks with your own
drawings.

36
 Appendix 2

Practical 2 - Lesson Plan

Microscopic observation of cells and tissues.

Context

A practical investigation set in the context of 9700 syllabus – Cell structure and
transport

Key aims of the lesson

This practical is designed to develop the skills of using a microscope and the
recording and interpretation of observations by producing biological drawings.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Use a compound light microscope
• Make clear and accurate plan and cellular drawings of biological tissue
• Be able to interpret structures seen through the microscope

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – Student worksheet given out for students to read in

previous preparation for the practical lesson. Students to look at examples of
lesson drawings of cells and tissues in their textbooks.

0-3 Introduction to the aims, intended outcomes and shape of the lesson
– teacher led oral presentation

3-5 Context – review the protocols for setting up and using a microscope
to observe slides

5 - 10 Introduction to method – Teacher briefly outlines method and

answers any student questions on procedure. Teacher emphasises
safety concerns regarding breaking slide by focussing down with the
objective lens.

10 - 25 Carrying out the practical – students carry out the practical work.

37
 Appendix 2

25 - 50 Obtain results – Students observe the plant tissue and produce clear
labelled diagrams as requested, then clear away apparatus as soon
as they have finished

50 - 60 Drawing together the threads – Teacher led discussion on the

manipulation and observational skills that have been developed as
well as discussion on results obtained. Teacher to go through the
mark scheme with the students and students compare their marks
and understand why marks were not awarded.

38
 Appendix 2

Practical 2 - Technical information

Microscopic observation of cells and tissues

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. 1 x prepared slide TS Lamium stem
2. 1 x microscope with suitable illumination and;
• high-power objective lens e.g. x40 (equal to 4mm or 1/6”)
• low-power objective lens e.g. x10 (equal to 16mm or 2/3”)
3. suitable white paper, HB (Medium hard) pencil and rubber
Safety Precautions.
No specific hazards have been identified in this practical, however a risk assessment
should be carried out as a matter of course.

39
 Appendix 2

Practical 3 - The identification of biological chemicals present in solutions

This practical focuses on making decisions about measurements and observations,
recording and presenting data and observations, interpretation, drawing conclusions
and suggesting improvements. You will also develop other assessed skills
throughout the practical.
Intended learning outcomes
By the end of this practical you should be able to:
• Decide what tests to carry out and what observations to make
• Use an appropriate means to record your observations, constructing any
tables before you make the observations
• Describe and summarise the key points of your observations
• Draw conclusions in terms of the presence or absence of different chemicals
in the solutions
• Suggest alternative strategies for identifying some of the materials

Safety information

You should wear eye protection throughout this practical.

Amylase is harmful, avoid contact with eyes and skin.

Benedict’s solution is harmful and dangerous to the environment.

1 mol dm–3 hydrochloric acid is harmful.

Background information
• Make sure that you know how to carry out Benedict’s test, what it is used for
and what the positive and negative results should be.
• Make sure that you know how to carry out biuret test, what it is used for and
what the positive and negative results should be.
• Think about how Benedict’s test and the enzyme amylase can be used to
confirm the presence of a polysaccharide such as starch.
• Think about how acid hydrolysis, neutralisation and Benedict’s test can be
used to confirm the presence of the non-reducing disaccharide, sucrose.

40
 Appendix 2

You will use the materials provided to identify the unknown materials in the solutions
A, B, C and D
• Read and think about the information above.
• The solutions A, B, C, D and E each contain only one of the following
materials, but not necessarily in this order
o A reducing sugar
o A non-reducing sugar
o A polysaccharide that can be hydrolysed by amylase
o Proteins including amylase
o No dissolved material
• You are also provided with materials for biuret test and for Benedict’s test,
as well as dilute hydrochloric acid, calcium hydrogen carbonate powder and
a waterbath at 35°C

Method
Preparations and making observations
1. You need to decide what tests to do and in what order so that it is possible
to use the amylase to test some of the other solutions.
2. Decide how you are going to record your observations so that it will be
absolutely clear what you did to which solutions, what you observed and
your interpretation of the observations.
3. Prepare a piece or pieces of paper in accordance with your decisions.
4. Make a risk assessment of your proposed methods and decide what
precautions to take to reduce the likelihood of an accident and to reduce the
damage any accidents might cause – ask your teacher to confirm that you
may go ahead with the tests.
5. Carry out the tests with full regard to safety, recording your observations and
interpretations.
6. Record the identity of the unknown solutions.

Write-up
• hand in your original laboratory records, including your methods,
observations and interpretations.
• suggest improvements to the method including some of the following:
• a simpler way of testing for the presence of starch,
• starch would also be hydrolysed by acid. Suggest a better order to do
your tests if this caused you difficulties, or a way of using amylase to
confirm that it is non-reducing sugar rather than starch that is present,
• if a solution contained a small amount of reducing sugar and also non-
reducing sugar, suggest how it might be possible to use repeated
benedict’s tests, filtering the precipitate out after each, to remove the
reducing sugar before testing for non-reducing sugar.

41
 Appendix 2

Practical 3 - Lesson Plan

The identification of biological chemicals present in solutions
Context
A practical investigation set in the context of 9700 syllabus – The identification of
biological chemicals present in solutions
Key aims of the lesson
This practical is designed to develop the skills of decision-making, observation,
interpretation and evaluation.
Intended learning outcomes
By the end of this practical the student should be able to:
• Decide what tests to carry out and what observations to make
• Use an appropriate means to record your observations, constructing any
tables before you make the observations
• Describe and summarise the key points of your observations
• Draw conclusions in terms of the presence or absence of different chemicals
in the solutions
• Suggest alternative strategies for identifying some of the materials
Resources required
White board or flipchart and suitable pens or blackboard and chalk
Practical materials specified on the Technical Information Sheet.
Copies of the student worksheets.
Planned activities
Timings/ Teacher/ Student Activities
minutes
End of Preparation – student worksheet given out for students to read in
previous preparation for the practical lesson. To ensure understanding of
lesson methods and their use as well as to consider decisions about which
observations to make and in what order, and the means of recording
0-3 Introduction to the aims, intended outcomes and shape of the lesson
– teacher led oral presentation
3-5 Context – review of biochemicals and biochemical tests - key points
written on board
5-8 Introduction to method – Teacher briefly reminds students of
methods of tests, demonstrating as necessary. Teacher answers any
student questions on procedure. Teacher emphasises safety concerns
with acidic and harmful solutions and heating of solutions
8 - 45 Carrying out the practical – students carry out the practical work and
record their observations and interpretations.
45 - 50 Clear up – Students clear away apparatus as soon as they have
finished

42
 Appendix 2

50 - 60 Drawing together the threads – Teacher led discussion on the skills

that have been developed as well as discussion on results obtained.
Teacher to emphasise importance of handing in original laboratory
records – (these can be collected in at the end of the lesson,
photocopied and given back to students if they are thought likely to
‘copy them up’). Practical write up to be completed in following lesson
or as homework activity.

Useful information
The most effective methodology is likely to be
• Test a small sample of each of the solutions with biuret test to identify the
protein including amylase – set aside solution B for later use (identified with
this test)
• Carry out Benedict’s test, heating the tubes gently to 90°C until one gives a
positive result, on small samples of each of solutions A, C, D and E to
identify which contains reducing sugar – set aside solution A (identified with
this test)
• Mix small samples from each of solutions C, D and E with equal volumes of
solution B and incubate at 35°C for a few minutes. Test sample from each
of the incubated tubes with benedict’s test to identify in which the
polysaccharide has been hydrolysed to give a reducing sugar – set aside
tube E (identified with this test)
• Mix small samples from tubes C and D with an equal volume of hydrochloric
acid and boil carefully for two minutes. Add a small excess of calcium
hydrogen carbonate (until it just stops effervescing) to neutralise the acid
and test with Benedict’s test to identify which contains non-reducing sugar
(C) and therefore which contains only water (D).
Summary of contents of the five solutions
solution material present in solution
A reducing sugar
B proteins including amylase
C non-reducing sugar
D no dissolved material
E polysaccharide that can be hydrolysed by amylase
• To improve the methodology, iodine solution would be a more elegant way
of confirming the presence of the polysaccharide.
• To detect non-reducing sugar in the presence of reducing sugar – react the
latter with excess Benedict’s and filter out the resulting precipitate. Re-test
to confirm the absence of reducing sugar. Now hydrolyse with acid,
neutralise and re-test.

43
 Appendix 2

Practical 3 - Technical information

The identification of biological chemicals present in solutions
The apparatus and materials required for this practical are listed below.
The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1 5 small beakers or other containers as follows
label on 20 cm3 of solution in beaker
beaker
A 1 mol dm-3 glucose solution made by dissolving 18 g of glucose in 80
cm3 of distilled water and making up to 100 cm3
B 10% egg white and 1% amylase solution made up by dissolving 10 cm3
of fresh egg white (or 1 g of dried egg white) and 1 g of amylase in 80
cm3 of cold distilled water, mixing until dissolved and making up to 100
cm3
C 0.5 mol dm-3 sucrose solution made by dissolving 17 g of sucrose in 80
cm3 of distilled water and making up to 100 cm3
D no dissolved material
E polysaccharide that can be hydrolysed by amylase

2 5 test tubes in a rack and a means of washing the tubes such as a sink and
running water
3 a glass marker, such as a wax pencil or a permanent OHP pen or small
labels and pencil
4 the usual materials that the students are used to using for biuret test,
labelled appropriately
5 the usual materials and heating arrangements that the students are used to
using for Benedict’s text, labelled appropriately
6 1 mol dm-3 hydrochloric acid in a small dropper bottle, labelled hydrochloric
acid
7 Sodium hydrogen carbonate powder in a small specimen tube with a
stopper and a spatula that fits in the tube to dispense it
8 A thermostatic waterbath or plastic trough containing water at about 35°C for
use as a waterbath
Safety Precautions/Risks

Amylase = H

Benedicts solution = H, N

Hydrochloric acid (1 mol dm–3) = H

A risk assessment should be carried out as a matter of course.

44
 Appendix 2

Practical 4 - Investigation of the carbohydrates metabolised by yeast

This practical focuses on making measurements and observations, recording and

presenting data, analysis, drawing conclusions and evaluating methods. You will also
develop other assessed skills throughout the practical.

Intended learning outcomes

By the end of this practical you should be able to:

• Experience relevant methods, analysis and conclusion.
• Describe and explain the relationship between temperature and membrane
permeability.
• Evaluate procedures

Safety Information

You should wear eye protection throughout this practical.

Methylene Blue is harmful. Avoid contact with eyes and skin. It will stain
skin or clothes.

Background information
• Yeast can metabolise carbohydrates under two different conditions. When
oxygen is present aerobic respiration occurs yielding a large amount of
energy for the organism and producing carbon dioxide & water as waste
products.
• However when oxygen is in short supply (anaerobic conditions) the yeast
will break down the carbohydrate into ethanol & carbon dioxide with a
much reduced energy output (alcoholic fermentation).
• Both of these forms of respiration in addition to most metabolic processes
are catalysed by specific enzymes.
• The process of how efficient the yeast is in metabolising different
carbohydrates can be monitored by observing the time taken for Methylene
Blue to be discoloured

In this experiment you will investigate the relative efficiency with which different
carbohydrates can be metabolised by yeast.

• Read the information above

• Identify and write down the dependent and independent variables
• Consider which type of carbohydrate (monosaccharide, disaccharide,
polysaccharide) will be metabolised by the yeast and why. Explain your
reasoning.
• Write down what you expect to happen as a hypothesis in which you make
specific predictions about which carbohydrates you might expect yeast to
metabolise.

45
 Appendix 2

• Identify any variables that should be controlled and outline how this should
be done

Method

Preparations and making observations

1. Label seven boiling tubes A - G.
2. In tube A place 5cm3 distilled water, in tube B 5cm3 glucose, in tube C
5cm3 fructose and continue with as many carbohydrates provided placing
each tube in a rack.
3. Into each tube add 3 drops of Methylene blue.
4. Add 5cm3 yeast solution to each tube noting the time.
5. Shake each tube to mix the contents and place back into the rack.
6. Do not disturb the tubes again but note the time taken for the blue colour to
disappear from each tube.

Write-up
• Record your results in a clear table ensuring units are put in headers
where possible.
• Represent the results of the experiment in a suitable chart to show type of
carbohydrate against the time taken for the blue colour to disappear.
• Explain your findings in terms of enzymes activity and carbohydrate
structure.
• Assess the reliability of the results obtained and suggest any modifications
you could make to improve the experiment

46
 Appendix 2

Practical 4 - Lesson Plan

Investigation of the different carbohydrates metabolised by yeast.

Context

A practical investigation set in the context of 9700 syllabus – Enzymes, biological

molecules, respiration.

Key aims of the lesson

This practical is designed to develop the skills of observation, analysis and

evaluation.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Experience relevant methods, analysis, conclusions and evaluation.
• Describe and explain the relationship between temperature and the
permeability of cell membranes.

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – Student worksheet given out for students to read in

previous preparation for the practical lesson. To consider identification of the
lesson variables, formulate a hypothesis and review previous learning on
cell membranes

0-3 Introduction to the aims, intended outcomes and shape of the

lesson – teacher led oral presentation

3-5 Context – review of enzyme controlled reactions, biological

molecules & respiration. Key points written on board

5-8 Introduction to method – Teacher briefly outlines method and

answers any student questions on procedure. Teacher emphasises
safety concerns with methylene blue

8 - 40 Carrying out the practical – students carry out the practical work.
Whilst they are waiting for the colour change to occur they can write
up the first part, identifying variables, hypothesis, results table.

47
 Appendix 2

40 - 50 Obtain results – Students enter results into table and clear away
apparatus as soon as they have finished

50 - 60 Drawing together the threads – Teacher led discussion on the

skills that have been developed as well as discussion on results
obtained. Practical write up to be completed in following lesson or as
homework activity

48
 Appendix 2

Useful information
Safety precautions:
• Methylene blue is harmful and can be a skin irritant. Safety glasses
should be worn. Additionally as it is a protein stain it will stain any
natural material. Please emphasise to students the importance of
safety when pipetting the methylene blue.
Discussion / evaluation points should include:
• Why should the tubes remain still after the initial mixing?
• What is being measured by the methylene blue discolouration (i.e. removal
of oxygen from the system by the aerobically respiring yeast) ?
• Suggest why some sugars are metabolised and others are not.
• Why was the yeast incubated for about 30 minutes before the experiment
started?
• What was the purpose of the tube with distilled water and yeast solution?
• Ensure that the students are aware of what type of organism that yeast
belongs to.
• Yeasts live in many different environments. Suggest why the following are
suitable places for yeast growth
a) fruit skin
b) Human body
• What precautions could be undertaken to ensure that all the tubes remained
at a constant temperature?
• for students unable to obtain a full set of results the following could be used
for analysis. Please note that other students results may not agree with
these ones.

Type of carbohydrate Time taken for blue colour to

disappear (minute:secs)

Glucose 6:15

Fructose 24:45

Galactose No change

Lactose No change

Maltose 25:30

Sucrose 8:40

Starch 42:00

49
 Appendix 2

Practical 4 - Technical information

The metabolism of different carbohydrates by yeast

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. 5cm3 of as many of the following carbohydrates as available. Each made up
to 5% concentration – Fructose, Galactose, Glucose, Lactose, Maltose,
Starch, Sucrose
2. 5cm3 distilled water
3. 7 boiling tubes (or as many as the number of carbohydrates available plus
control)
4. Methylene blue, 3 drops per sample
5. 5cm3 yeast solution (prepared in advance) per sugar used
6. test tube rack
7. Timer
8. Safety glasses

Additionally each student will require access to a sink & running water.

The yeast should be prepared according to local conditions so that it is activated and
ready for use.
Safety Precautions/Risks.

Methylene blue = H

A risk assessment should be carried out as a matter of course.

50
 Appendix 2

Practical 5 - The effect of pH on enzymes

This practical focuses on making measurements and observations, recording and

presenting data, analysis, drawing conclusions and evaluating methods. You will also
develop other assessed skills throughout the practical.

Intended learning outcomes

By the end of this practical you should be able to:

• Identify dependent and independent variables
• Make a hypothesis and express this in words and graphically
• Experience relevant methods, analysis and conclusion.
• Describe and explain the relationship between ph and enzyme activity
• Evaluate procedures

Safety Information

You should wear eye protection throughout this practical.

Hydrogen peroxide is corrosive. Avoid contact with eyes or skin. It will

bleach skin or clothes.

Citric acid is harmful.

Background information
• Most enzymes have an optimum pH near to 7 (the pH found inside most
cells)
• pH is the measurement of the concentration of Hydrogen ions (H+)
• Hydrogen ions will affect the hydrogen and ionic bonds within the enzyme
• If these bonds are changed the three dimensional shape is changed
altering the shape of the active site
• When an enzymes shape is altered it becomes denatured
• Potatoes are a good source of catalase

You will investigate the effect of pH on the enzyme catalase as it breaks down toxic
Hydrogen peroxide, a by-product of some biochemical reactions, into water and
oxygen.
• Read the information above
• Identify and write down the dependent and independent variables
• Write down a hypothesis
• Draw a sketch graph to show what you think will happen

51
 Appendix 2

• Identify any variables that should be controlled and outline how this should
be done
• What would be the best method for collecting the oxygen produced?
• A graph of pH against rate of activity will be produced after the practical.
Make sure you know how to calculate rate.
• Know what a buffer solution is and what it does.

Method

Preparations and making observations

1. Use a cork borer to cut cylinders of fresh potato tissue. You will require a
piece (pieces) between 6 – 7cm in length. Place on a tile and cut into at
least 60 discs, each 1mm wide.
2. Place all the discs in a small beaker of water.
3. Set up the equipment as follows. Clamp a boiling tube to a stand and
carefully insert the manometer (with fluid) into a rubber bung.
4. Using a syringe or small measuring cylinder place 5cm3 of buffer solution
pH3 into the boiling tube.
5. Carefully add 10 of the potato discs followed by 5cm3 of hydrogen peroxide
solution.
6. Place the bung back into the boiling tube as quickly as possible.
7. Start the stop watch and time how long it takes for the manometer fluid to
rise by 5cm. (Mark start point on tube and measure 5cm)
8. Carefully agitate the tube to make sure that the potato discs do not stick
together.
9. Wash out the boiling tube and repeat the experiment using a different pH
buffer and ten new potato discs each time.
10. Make sure you use a clean syringe / measuring cylinder with each different
buffer solution.
11. If time allows repeat the procedure for increased reliability.

Write-up
• Record your results in a clear table ensuring units are put in headers.
• If replicate results not done obtain a set of mean readings by using other
class members results.
• Calculate the rate of reaction.
• Plot a graph of pH against rate.
• Explain your findings using your knowledge of enzymes.
• Assess the reliability of the results obtained and suggest any modifications
you could make to improve the experiment
• How could you measure the volume of gas produced by this method and
by altering the method

52
 Appendix 2

Practical 5 - Lesson Plan

The effect of pH on enzyme activity

Context

A practical investigation set in the context of 9700 syllabus – The effect of pH on

enzyme activity

Key aims of the lesson

This practical is designed to develop the skills of observation, analysis and

evaluation.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Experience relevant methods, analysis, conclusions and evaluation.
• Describe and explain the relationship between pH and enzyme activity.

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – 2 page student worksheet given out for students to

previous read in preparation for the practical lesson. To consider identification
lesson of the variables, formulate a hypothesis and review previous learning
on cell membranes

0-3 Introduction to the aims, intended outcomes and shape of the

lesson – teacher led oral presentation

3-5 Context – review of enzymes, key points written on board

5-8 Introduction to method – Teacher briefly outlines method and

answers any student questions on procedure. Teacher emphasises
safety concerns with cork borer (or sharp knife) and hydrogen
peroxide

8 - 45 Carrying out the practical – students carry out the practical work..

45 - 50 Obtain results – Students enter results into table and clear away
apparatus as soon as they have finished

53
 Appendix 2

50 - 60 Drawing together the threads – Teacher led discussion on the

skills that have been developed as well as discussion on results
obtained. Practical write up to be completed in following lesson or as
homework activity

Useful information
Discussion / evaluation points should include:
• explanation of the shape of the graph
• consistency of the enzyme within the potato, age of potato
• possible problems with the method e.g. Lack of temperature control in
reaction tube, loss of gas before bung inserted
• for students unable to obtain a full set of results the following could be used
for analysis

pH Time taken for manometer to

move 5cm (minutes)

3 10

4 8

5 6

6 4

7 2

8 5

9 8

10 10

54
 Appendix 2

Practical 5 - Technical information

The effect of pH on enzymes

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. fresh potato (each student requires a core (cores) approximately 10cm in
length). (More if experiment is to be repeated)
2. 1 boiling tube
3. Single bore rubber bung
4. cork borer
5. white tile
6. scalpel
7. small beaker
8. 2 x 10cm3 graduated pipette or measuring cylinder or syringe
9. Manometer tube (3mm diameter)
10. Stop watch
11. Forceps
12. 20 volume Hydrogen peroxide
13. Range of buffers (pH 3 – 8).

Additionally each student will require access to a sink and running water.

Commercial buffer tablets are available from most chemical wholesalers, however it
is possible to make up buffer solutions in the laboratory. (Details from, Laboratory
Manual for Schools. Heinemann. 1977)

Sodium hydrogen phosphate/citric acid buffer – range pH 3.0 – 8.0.

To make up 100cm3 of buffer use 0.1M Citric Acid & 0.2M Sodium hydrogen
phosphate in the following proportions:
Citric acid / cm3 Sodium hydrogen
pH
phosphate / cm3

3.0 79.45 20.55

4.0 61.45 38.55

5.0 48.50 51.50

6.0 36.85 63.15

7.0 17.65 82.35

8.0 2.75 97.25

55
 Appendix 2

Safety Precautions/Risks.

Hydrogen peroxide = C

Citric acid = H

A risk assessment should be carried out as a matter of course.

56
 Appendix 2

Practical 6 - The effect of inhibitors on enzyme activity

This practical focuses on making measurements and observations, recording and

presenting data, analysis, drawing conclusions and evaluating methods. You will also
develop other assessed skills throughout the practical.

Intended learning outcomes

By the end of this practical you should be able to:

• Experience relevant methods, analysis and conclusion.
• Describe and explain the effect of a non competitive inhibitor on enzyme
activity.
• Evaluate procedures.

Safety Information

You should wear eye protection throughout this practical.

Amylase is harmful. Avoid contact with eyes or skin.

Iodine solution is harmful. Avoid contact with eyes or skin. It will stain skin
or clothes.

Background information
• A non competitive inhibitor binds to a part of the enzyme away from the
active site
• The shape of the enzyme is changed, thus changing the shape of the
active site
• No enzyme-substrate complexes can be formed, hence no product
produced
• Increasing the amount of substrate does not overcome the effect of this
type of inhibitor

You will investigate the effect of increasing the amount of Lead nitrate on the
hydrolysis of starch by the enzyme amylase.
• Read the information above
• Identify and write down the dependent and independent variables
• Write down a hypothesis
• Draw a sketch graph to show what you think will happen
• Identify any variables that should be controlled and outline how this should
be done

57
 Appendix 2

A colorimeter should be used to compare the colours of the solutions obtained after a
given time.

Method

Preparations and making observations

1. Set up a thermostatically controlled water bath set at 40°C
2. Label six boiling tubes A – F and place in a test tube rack
3. Add 10cm3 starch solution to each tube
4. Add the following quantities of Lead nitrate to 5cm3 distilled water in six
test tubes labelled 1 – 6 : 0g, 0.1g, 0.2g, 0.3g, 0.4g, 0.5g and shake to
ensure it dissolves
5. Pour contents of tube 1 into boiling tube A, tube 2 into boiling tube B etc.
6. Add 5cm3 amylase solution to each of the boiling tubes, agitate well and
start the timer after placing the tubes into the water bath
7. Allow the reaction to proceed for 20 minutes
8. During this time adjust the colorimeter using a solution of 1cm3 iodine
solution in a boiling tube containing 10cm3 starch solution and 5cm3
distilled water. Set the colorimeter to 0% transmission with this solution
9. After 20 minutes add 1cm3 iodine solution to each tube
10. Test each of the tubes A – F in the colorimeter noting down the
absorbance for each tube and record in a table. (Note for centres without
access to a colorimeter the method could be adapted to using a spotting
tile with one drop of iodine in each well. At intervals of one minute a drop of
the reaction mixture is placed on the tile and the time taken for the black
colour to disappear noted.)

Write-up
• Record your results in a clear table ensuring units are put in headers
where possible.
• Plot a graph of transmission against mass of lead nitrate added. (If
experiment done using spotting tile method a graph of rate of reaction
against mass of lead nitrate added should be drawn)
• Explain your findings using your knowledge of enzymes and inhibitors.
• Assess the reliability of the results obtained and suggest any modifications
you could make to improve the experiment.
• Why was it necessary to control the temperature of the reaction?
• What further experiments could be done to investigate non-competitive
inhibitors?
• What are the advantages of repeating an experiment?
• Suggest why the iodine solution was not added at the same time as the
other solutions

58
 Appendix 2

Practical 6 - Lesson Plan

The effect of inhibitors on enzyme activity

Context

A practical investigation set in the context of 9700 syllabus – Enzymes and enzyme
inhibitors

Key aims of the lesson

This practical is designed to develop the skills of observation, analysis and

evaluation.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Experience relevant methods, analysis, conclusions and evaluation.
• Describe and explain the effect of a non competitive inhibitor

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – Student worksheet given out for students to read in

previous preparation for the practical lesson. To consider identification of the
lesson variables, formulate a hypothesis and review previous learning on
enzymes.

0-3 Introduction to the aims, intended outcomes and shape of the

lesson – teacher led oral presentation

3-5 Context – review of enzyme action,

5-8 Introduction to method – Teacher briefly outlines method and

answers any student questions on procedure. Teacher emphasises
safety concerns with the use of chemicals.

8 - 40 Carrying out the practical – students carry out the practical work.
Whilst they are waiting for the 20 minute period they can write up the
first part, identifying variables, hypothesis, results table. Teacher to
demonstrate the use of colorimeter to those students unfamiliar with
this piece of equipment.

59
 Appendix 2

40 - 50 Obtain results – Students enter results into table and clear away
apparatus as soon as they have finished

50 - 60 Drawing together the threads – Teacher led discussion on the skills

that have been developed as well as discussion on results obtained.
Practical write up to be completed in flowing lesson or as homework
activity

Useful information
• For centres without access to a colorimeter the practical results will not be
as accurate but will still be of an objective nature. The pupils could evaluate
to suggest improvements to include a more objective measurement.
• If the centre does not have access to thermostatically controlled water
baths, manually controlled ones could be substituted or left out altogether
but the need for temperature control needs to be discussed in the
evaluation.
Other Discussion / evaluation points should include:
• explanation of the shape of the graph
• the differences between competitive and non competitive inhibitors needs to
be emphasised
• for students unable to obtain a full set of results the following could be used
for analysis

Mass of Lead nitrate Transmission

/g / arbitrary units

0.0 85

0.1 52

0.2 37

0.3 27

0.4 23

0.5 21

60
 Appendix 2

Practical 6 - Technical information

The effect of inhibitors on enzyme activity

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. 6 boiling tubes.
2. 6 test tubes
3. test tube rack
4. labels/marker pen
5. 1% starch solution – allow 100cm3 per repeat
6. Amylase solution 1% - allow 50cm3 per repeat
7. Iodine solution – allow 10cm3 per repeat
8. Distilled water – 50cm3 per repeat
9. 10cm3 graduated pipette / measuring cylinder / syringe
10. Stopclock
11. colorimeter cuvettes

Additionally each student will require access to a colorimeter, thermostatically

controlled water bath, sink & running water.

If using spotting tile method also add 1 x spotting tile and glass rods.
Safety Precautions/Risks.

Amylase = H

Iodine solution = H

A risk assessment should be carried out as a matter of course.

61
 Appendix 2

Practical 7 - The effect of temperature on membrane permeability in beetroot

This practical focuses on making measurements and observations, recording and

presenting data, analysis, drawing conclusions and evaluating methods. You will also
develop other assessed skills throughout the practical.

Intended learning outcomes

By the end of this practical you should be able to:

• Experience relevant methods, analysis and conclusion.
• Describe and explain the relationship between temperature and membrane
permeability.
• Evaluate procedures

Safety Information

You should wear eye protection throughout this practical.

There are no particular hazards in this practical, however you must follow your
laboratory rules.

Background information
• The colour of beetroot is due to the presence of a red pigment called
anthocyanin
• The cell membrane is mainly made up of two types of molecules,
phospholipids and proteins scattered around in the membrane.
• The membrane is partially permeable
• Protein structure is denatured at high temperatures

You will investigate the effect of temperature on the permeability of the cell
membrane in beetroot.

• Read the information above

• Identify and write down the dependent and independent variables
• Write down a hypothesis
• Draw a sketch graph to show what you think will happen
• Identify any variables that should be controlled and outline how this should
be done

A colorimeter should be used to compare the colours of anthocyanin solutions

obtained.

62
 Appendix 2

Method

Preparations and making observations

1. Use a cork borer to cut cylinders of fresh beetroot tissue. Place on a tile
and cut into 30 discs, each 3mm wide.
2. Place all the discs in a small beaker and wash under a running tap for at
least five minutes.
3. Label six test tubes – 30°C, 40°C, 50°C, 60°C, 70°C, 80°C.
4. Add 10cm3 cold distilled / de-ionised water to each tube.
5. Set up a water bath using a large beaker, tripod, gauze and Bunsen
burner.
6. Heat the water gently until a temperature of 80oc is reached then remove
heat source.
7. Take five of the beetroot discs and impale on a mounted needle with space
between each disc.
8. Immerse the discs in the water bath for exactly one minute, then remove
and carefully push the discs into the test tube labelled 80°C and set aside.
9. Reduce the temperature of the water bath to 70°C and take a second set
of five discs and repeat the process of immersion for one minute followed
by putting them into the next tube.
10. Continue the process for each of the temperatures.
11. After the discs have stood for thirty minutes shake the tubes and pour this
liquid into a cuvette.
12. Fill a second cuvette with distilled water.
13. Place a blue filter into the colorimeter and use the distilled water to zero
the machine adjusting the pointer to zero absorbance.
14. Measure the colour density of the 70°C solution.
15. Wash out the cuvette and repeat the procedure to record the light
absorbance for each of the temperatures.

Write-up
• Record your results in a clear table ensuring units are put in headers
where possible.
• Plot a graph of relative concentration of pigment against temperature.
• Explain your findings using your knowledge of cell membranes.
• Assess the reliability of the results obtained and suggest any modifications
you could make to improve the experiment

63
 Appendix 2

Practical 7 - Lesson Plan

The effect of temperature on the permeability of cell membranes

Context

A practical investigation set in the context of 9700 syllabus – cell membranes and the
effect of temperature.

Key aims of the lesson

This practical is designed to develop the skills of observation, analysis and

evaluation.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Experience relevant methods, analysis, conclusions and evaluation.
• Describe and explain the relationship between temperature and the
permeability of cell membranes.

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – 2 page student worksheet given out for students to

previous read in preparation for the practical lesson. To consider identification
lesson of the variables, formulate a hypothesis and review previous learning
on cell membranes

0-3 Introduction to the aims, intended outcomes and shape of the

lesson – teacher led oral presentation

3-5 Context – review of cell membranes, key points written on board

5-8 Introduction to method – Teacher briefly outlines method and

answers any student questions on procedure. Teacher emphasises
safety concerns with cork borer (or sharp knife) and water baths

8 - 40 Carrying out the practical – students carry out the practical work.
Whilst they are waiting for the 30 minute period they can write up the
first part, identifying variables, hypothesis, results table. Teacher to
demonstrate the use of colorimeter to those students unfamiliar with
this piece of equipment.

64
 Appendix 2

40 - 50 Obtain results – Students enter results into table and clear away
apparatus as soon as they have finished

50 - 60 Drawing together the threads – Teacher led discussion on the skills

that have been developed as well as discussion on results obtained.
Practical write up to be completed in flowing lesson or as homework
activity

Useful information
• For centres without access to a colorimeter the practical results will only be
of a subjective nature. However the principles of denaturation of the proteins
in the membrane can still be discussed and the pupils could evaluate to
suggest improvements to include a more objective measurement.
• If the centre has access to a number of thermostatically controlled water
baths these could be substituted for the individual ones suggested in the
method.
Discussion / evaluation points should include:
• explanation of the shape of the graph
• consistency of the pigment within the beetroot
• why the discs were washed before heating
• possible problems with the method e.g. impaling onto a mounted needle
• for students unable to obtain a full set of results the following could be used
for analysis

Temperature Absorbance
/ °C / arbitrary units

30 0.12

40 0.16

50 0.29

60 0.83

70 1.62

80 1.41

65
 Appendix 2

Practical 7 - Technical information

The permeability of beetroot cell membrane

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. fresh beetroot (each student requires a core approximately 10cm in length).
2. 6 test tubes
3. test tube rack
4. cork borer
5. white tile
6. scalpel
7. small beaker
8. 10cm3 graduated pipette or measuring cylinder
9. mounted needle
10. large beaker
11. thermometer
12. Bunsen burner
13. tripod
14. heat proof mat
15. gauze
16. colorimeter cuvettes (2)

Additionally each student will require access to a sink, running water and a
colorimeter.
Safety Precautions/Risks.
No specific hazards identified.

A risk assessment should be carried out as a matter of course.

66
 Appendix 2

Practical 8 - Broad bean root tip squash

This practical focuses on setting up and manipulating apparatus and making and
recording observations. Further skills can be developed using additional information
after the practical has been completed.

Intended learning outcomes

By the end of this practical you should be able to:

• Experience simple techniques to observe mitosis in cells in the root tip of
broad beans
• Further your knowledge about mitosis
• Interpret and record observations

Safety Information

You should wear eye protection throughout this practical.

Acetic orcein is corrosive. Avoid contact with eyes or skin. It will stain skin
and clothes.

Background information
• The role of chromosomes is to store information in the DNA coding and be
able to replicate by cell division.
• Mitosis is a type of cell division found in somatic cells, that produces
diploid cells.
• Chromosomes are only visible when the cell is dividing. They can be made
more visible using appropriate staining techniques.
• During mitosis each chromosomes separates into two chromatids.
• Each chromatid is pulled to opposite ends of the spindle prior to the cell
dividing into two.
• The different stages of cell division are called interphase, prophase,
metaphase, anaphase, telophase and back to interphase and are
characterised by the position of the chromatids.
• During interphase the DNA of the chromatids replicates so that at
prophase, whole chromosomes are visible once more.
• Mitosis can be observed in the meristematic tissues found in the apical
meristem of actively growing roots of plants such as broad bean.

You will investigate how root tip tissue can be prepared and stained so that
chromosomes in cells undergoing mitosis can be viewed through a microscope. You
will then identify the various stages of cell division.

67
 Appendix 2

• Read the information above.

• Read your textbook and look carefully at the diagrams and photographs on
the section concerning mitosis.
• Write down the key features that will enable you to identify root tip cells at
each of the different stages of mitotic cell division.

Method

Preparation
1. You have been provided with a germinated seedling of a broad bean or
similar.
2. Using a sharp knife or scalpel, carefully cut of 5mm from the apical tip of
five lateral roots.
3. Place the root tips into a test-tube containing acetic orcein stain.

Warning! Acetic orcein is an acidic stain that turns protein purple. Any stain that gets
on skin or natural clothing will stain the material purple.
4. Warm the acetic orcein and root tips in a hot water bath but do NOT allow
the solution to boil. The stain should be kept so that it is gently steaming
for at least five minutes.
5. Using a mounted needle or similar, carefully remove one of the root tips.
The root tips will now be very soft and easily damaged so do not remove
them with forceps.
6. Leave the remaining four root tips in the hot stain.
7. Place the root tip onto a clean microscope slide.
8. Using a sharp knife cut it in half transversely so that each length is
approximately 2.5mm.
9. Discard the half that is furthest away from the root tip.
10. Using a pipette, carefully add two or three drops of acetic orcein stain to
the root tip on the slide.
11. Using a mounted needle or similar gently break the tissue of the root tip
apart.
12. Carefully lower a cover slip over the root tip and using the blunt end of
your mounted needle or a pencil, gently push the cover slip down onto the
slide so that the root tip is squashed. Care is needed to ensure that as few
air bubbles are present as possible.
13. Gently blot any excess stain from the slide.
14. Examine the slide using a microscope for the different stages in mitosis.
15. If insufficient staining has taken place, the process may be repeated with
the other four root tips that have been left in the hot stain.

Making observations
1. Initially observe the tissue using the low power of your microscope. This
will enable you to find areas of cells where the nuclear material is clearly
visible and undergoing mitosis.

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2. Using the high power of your microscope examine individual cells at

various stages of division.
3. Using the high power of you microscope make large, labelled drawings of
each of the stages of cell division.

Write-up
• Write up the method and answering the following questions
1. Suggest why the tissue was heated when placed in the acetic orcein stain.
2. Explain how you placed the coverslip on the slide in order to reduce the
number of air bubbles.
3. What was the diploid number of chromosomes in the cells of the root tip
that you examined?
4. Explain why the haploid number is always even.

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 Appendix 2

Practical 8 - Lesson Plan

Broad bean root tip squash

Context

A practical investigation set in the context of 9700 syllabus – Cell and nuclear
division

Key aims of the lesson

This practical is designed to develop the skills of manipulation of apparatus and

observation, and the recording and interpretation of observations.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Experience relevant methods.
• Describe and explain the reasons behind the methods.
• Extend knowledge on the structure and function of mitosis

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – Student worksheet given out for students to read in preparation for
previous the practical lesson. Students to read and look at drawings and photographs of
lesson mitosis in their textbooks.

0-3 Introduction to the aims, intended outcomes and shape of the lesson – teacher
led oral presentation

3-5 Context – review the sequence of mitosis and determine the key features of each
of the stages with key points written on board

5 - 10 Introduction to method – Teacher briefly outlines method and answers any

student questions on procedure. Teacher emphasises safety concerns when
using hot acetic orcein stain

10 - 25 Carrying out the practical – students carry out the practical work.

25 - 50 Obtain results – Students observe mitosis and produce clear labelled diagrams
of each stage, then clear away apparatus as soon as they have finished

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 Appendix 2

50 - 60 Drawing together the threads – Teacher led discussion on the manipulation and
observational skills that have been developed as well as discussion on results
obtained. Practical write up to be completed in following lesson or as homework
activity

Useful information
• Other plant root tips can be substituted for broad bean, such as other types
of bean, onions or sunflowers.
• The intensity of the stain depends upon how long the root tips are left in the
hot acetic orcein stain.
Discussion / evaluation points should include:
• explanation of the methods used and how to successfully produce a
mounted slide
• possible problems with the method such as the degree of staining, sufficient
squashing of tissue and microscope technique

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 Appendix 2

Practical 8 - Technical information

Broad bean root tip squash

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. 1 x actively growing broad bean seedling such that at least five lateral roots
have developed. Broad bean may be substituted with any other bean, or
onion or sunflower.
2. sharp knife or scalpel
3. 1 x test-tube and hot water bath that can be maintained at approximately
90°C
4. 1 x microscope slide and cover slip
5. 1 x mounted needle or similar
6. 5cm3 of acetic orcein stain. If not previously prepared, the acetic orcein
should be mixed with 1M HCl in the proportions of ten parts stain to one part
acid.
7. Access to microscope with both low and high power objective lenses
Safety Precautions/Risks.

Acetic orcein = C

A risk assessment should be carried out as a matter of course.

NOTE

Acetic orcein is an acidic stain that turns protein purple. Any stain that gets on
skin or natural clothing will stain the material purple.

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 Appendix 2

Practical 9 - The extraction of DNA from onions

This practical focuses on setting up and manipulating apparatus and making

observations. Further skills can be developed using additional information after the
practical has been completed.

Intended learning outcomes

By the end of this practical you should be able to:

• Experience simple techniques to extract DNA from living material.
• Further your knowledge about the structure of DNA

Safety Information

You should wear eye protection throughout this practical.

Protease is harmful. Avoid contact with eyes or skin.

Ethanol is highly flammable. There should be no flames in the same room.

Background information
• DNA is a polymer made up of monomers called nucleotides
• A gene is a set of coded instructions made up of a particular order of
nucleotides
• A nucleotide consists of three parts:
i) a pentose sugar
ii) a nitrogen containing base
iii) a phosphate group
• DNA molecule is a double helix held together by hydrogen bonds between
the complementary base pairs

You will investigate how DNA can be extracted from living material such as onion.

• Read the information above

• Read your text book and notes on DNA

Method

Preparations and making observations

1. Prepare the degrading mixture by adding 3g of sodium chloride (table salt)
to 10cm3 liquid detergent (washing up liquid). Make this up to 100cm3 with
distilled water and stir well to ensure the salt has dissolved.
2. The onion should be chopped into small pieces and added to the detergent

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 Appendix 2

solution and stir.

3. Place the beaker into a water bath maintained at 60oc for fifteen minutes.
4. Immediately cool down the onion mixture in an ice cold water bath for five
minutes again stirring frequently.
5. Blend using a food blender for no more than five seconds.
6. Using coffee filter paper (laboratory filter paper not coarse enough) filter
the mixture into a new beaker.
7. Once you have obtained enough liquid pour 10cm3 into a boiling tube and
add 2 – 3 drops of a protease enzyme and shake the tube to mix the
contents well.
8. Pour 10cm3 ice cold ethanol slowly into the boiling tube and place the tube
into rack for about five minutes.
9. DNA should appear where the two liquids meet.

Write-up
• Write up the method and answering the following questions
1. What effect would the washing up liquid (detergent) have on the cell
membranes?
2. Why was the beaker placed in a hot water bath for 15 minutes and then
immediately cooled?
3. Why was the mixture blended, but only for 5 seconds?
4. What type of enzyme would now be needed to separate the DNA into
smaller pieces?

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 Appendix 2

Practical 9 - Lesson Plan

The extraction of DNA from onions

Context

A practical investigation set in the context of 9700 syllabus –

Key aims of the lesson

This practical is designed to develop the skills of observation and manipulation of

apparatus.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Experience relevant methods.
• Describe and explain the reasons behind the methods.
• Extend knowledge on the structure and function of DNA.

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – Student worksheet given out for students to read in

previous preparation for the practical lesson. To consider the structure of
lesson DNA and reinforce previous learning

0-3 Introduction to the aims, intended outcomes and shape of the

lesson – teacher led oral presentation

3-5 Context – review of DNA structure, key points written on board

5-8 Introduction to method – Teacher briefly outlines method and

answers any student questions on procedure. Teacher emphasises
safety concerns with sharp knives and use of ethanol

8 - 45 Carrying out the practical – students carry out the practical work.
Whilst they are waiting for the 15 minute period they can write up the
first part of the method and consider the questions.

40 - 50 Obtain results – Students observe DNA produced then clear away

apparatus as soon as they have finished

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 Appendix 2

50 - 60 Drawing together the threads – Teacher led discussion on the

skills that have been developed as well as discussion on results
obtained. Practical write up to be completed in following lesson or as
homework activity

Useful information
• Other vegetables/fruit can be substituted for onions, however mixed results
are often obtained.
Discussion / evaluation points should include:
• explanation of the methods used
• possible problems with the method

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 Appendix 2

Practical 9 - Technical information

Extraction of DNA from onions

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. fresh onion, approximately tennis ball sized
2. sharp knife
3. chopping board
4. 2 x 250cm3 beakers
5. 1 x 400cm3 beaker or jug (for the ice)
6. 3g salt
7. 10cm3 washing up liquid
8. 90cm3 distilled water
9. (Thermostatically controlled) water bath at 60°C.
10. Supply of ice
11. Food blender (household domestic one is ideal)
12. Coffee filter paper
13. Funnel
14. Boiling tube
15. 2-3 drops of protease enzyme, such as neutrase ®
16. 10cm3 ice cold ethanol

SAFETY NOTE

The ethanol must be ice cold, this involves leaving it overnight in a freezer. It is
essential that it is placed in a sealed, vapour tight plastic bottle. If this is not possible
put the ethanol in a sealed container in an ice bath for several hours before the
practical is due to start.
Safety Precautions/Risks.

Protease = H

Ethanol = F

A risk assessment should be carried out as a matter of course.

77
 Appendix 2

Practical 10 - The effect of wind speed on the rate of transpiration in a leafy

shoot

This practical focuses on making measurements and observations, recording and

presenting data, analysis, drawing conclusions and evaluating methods. You will also
develop other assessed skills throughout the practical.

Intended learning outcomes

By the end of this practical you should be able to:

• Identify dependent and independent variables
• Make a hypothesis and express this in words and graphically
• Identify the variables that should be controlled
• Experience relevant methods, analysis and conclusion.
• Describe and explain the relationship between transpiration and wind speed
• Evaluate procedures

Safety Information

You should wear eye protection throughout this practical.

There are no particular hazards in this practical, however you must follow your
laboratory rules.

Background information
• Transpiration is the movement of water through plants, from the roots
where it is absorbed by osmosis, to the leaves where it is lost by
evaporation.
• Water leaves a plant’s leaves through stomata, the aperture of which is
controlled by guard cells.
• Most plants open their stomata during the day and close them at night.
• Plants may close their stomata when stressed by losing too much water.
• The real purpose of stomata is to absorb carbon dioxide and release
oxygen from photosynthesis without losing too much water.
• The evaporation of water is affected by wind speed, temperature, humidity
and atmospheric pressure.

You will investigate how wind speed affects the rate of transpiration from a leafy
shoot by using a potometer.

• Read the information above

• Identify and write down the dependent and independent variables
• Write down a hypothesis
• Draw a sketch graph to show what you think will happen

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 Appendix 2

• Identify any variables that should be controlled and outline how this should
be done
• What would be the best method for setting up the potometer?
• Plot a graph of the distance of the fan from the shoot, against the rate of
water movement in the potometer, after the practical. Make sure you know
how to calculate rate.

Wind speed in this case is varied by moving the fan to fixed distances from the leafy
shoot. Although the wind speed from the fan will not accurately follow the inverse -
square law, you would be well advised to understand how increasing the distance of
the fan from the shoot, may affect the wind speed.

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 Appendix 2

Method

Preparations

The apparatus should be assembled as shown in the following diagram.

1. Attach the rubber tubing, the capillary tube and the water reservoir to the T
piece.
2. Fill the reservoir, capillary tube and rubber tubing with water. This can be
done by placing them under water and gently squeezing the rubber tubing
until all the air has been removed.
3. Leave the apparatus under water.
4. Cut a fresh leafy shoot with a sharp knife and immediately place the cut
end under water.
5. Carefully attach the cut end of the shoot to the rubber tubing. This should
be done with the cut end only under water.
6. Close the tap on the water reservoir.
7. Remove the apparatus from the water and attached to a clamp stand or
support.
8. Place a mm scale behind the capillary tube.
9. Place a fan at a set distance from the leafy shoot. Do NOT switch on.

Making observations
1. Note the position of the air bubble in the capillary tube. It may be very
close to then end of the tube.
2. Record the time taken for the air bubble to move a set distance along the
tube. You will have to determine this distance base on the speed of the
bubble. If the bubble is moving quickly the distance will need to be larger
than if it is moving slowly.
3. Reset the air bubble to then end of the capillary tube by carefully opening
the tap on the water reservoir.

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 Appendix 2

4. Turn on the fan and repeat the procedure.

5. Reset the apparatus and move the fan to another distance.
6. Repeat the procedure with the fan at at least five differetn distances.
You are advised to start with the fan at the furthest distance and gradually move it
towards the leafy shoot.

Calculations
1. Calculate the rate of movement using 1/time taken for the air bubble to
travel a set distance.
2. Record the rate of travel for each distance in the class result table on the
board or flip chart.
3. When all of the results have been recorded in the class results table,
calculate the mean rate of movement for each distance.
4. (Optional) – calculate the standard error for each distance.

Write up

• Record your results in a clear table ensuring units are put in headers.
• Plot a graph to show the mean rate of movement for each distance.
• (Optional – add error bars to your graph)
• Make an evaluation considering:
• the limitations of the method used,
• anomalous values if any,
• replication and range of values of independent variable,
• the confidence with which the conclusions should be drawn.
• Draw conclusions considering:
• detailed description of the features of the results,
• whether your results agree or contradict your hypothesis,
• a scientific explanation of your results and conclusions,
• any modifications you could make to improve the experiment.

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 Appendix 2

Practical 10 - Lesson Plan

The effect of wind speed on the rate of transpiration in a leafy shoot.

Context

A practical investigation set in the context of 9700 syllabus – Investigate

experimentally the factors that affect transpiration rate.

Key aims of the lesson

This practical is designed to develop the skills of planning, observation, analysis and
evaluation.

Intended learning outcomes

By the end of the practical and the write-up the student should be able to:
• make a hypothesis and express this in words and graphically
• identify the dependant and independent variables
• identify the variables that should be controlled
• experience relevant methods, analysis, conclusions and evaluation
• describe and explain the relationship between wind speed and the rate of
transpiration in a leafy shoot.

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 Appendix 2

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities (timings can be altered to suit shorter or longer lessons)

Timings/ Teacher/ Student Activities

minutes

End of Preparation – 2 page student worksheet given out for students to

previous read in preparation for the practical lesson. To consider identification
lesson of the variables, formulate a hypothesis and review previous learning
on water movement through plants.

0-3 Introduction to the aims, intended outcomes and shape of the lesson
– teacher led oral presentation

3-5 Context – review transpiration in plants.

Teacher led questioning, student responses / discussion regarding the
factors that can affect the rate of transpiration and how they might be
measured.

5 - 10 Introduction to method – Teacher briefly demonstrates the

procedure for setting up the potometer and explains the importance of
not having any air in the apparatus until the introduction of the air
bubble. Teacher emphasises safety concerns with the sharp knife.

10 - 40 Carrying out the practical – students carry out the practical work..

40 - 50 Obtain results – Students enter results into table and clear away
apparatus as soon as they have finished

50 - 60 Drawing together the threads – Teacher led discussion on the skills

that have been developed as well as discussion on results obtained.
Practical write up to be completed in following lesson or as homework
activity to include identification of variables, the hypothesis, results
and graphs along with a full detailed write of the experiment and an
explanation of how the experiment could be extended.

Useful information
• The evaporation of water from a leaf is affected by wind speed, temperature,
humidity and air pressure, the first three having the most significant affect.
• Increased temperature increases the kinetic energy of the water molecules
thus increasing the rate of evaporation.
• Increased wind speed blows away evaporated molecules form around the
opening of the stomata thus maintaining a greater diffusion gradient for the
water molecules.

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 Appendix 2

• Increased humidity lowers the concentration gradient and thus slows down
the rate of evaporation.
• Reduced air pressure increases the rate of evaporation.
• Factors such as light are affected by the inverse square law where doubling
the distance reduces the light intensity by a factor of four. Although this does
not hold true for wind speed, you should be aware that doubling the distance
of the fan from the leafy shoot, will not necessarily mean that the wind speed
is reduced by half.
• Possible variables to control include temperature and humidity.
For students unable to obtain accurate data, the following table of results may be
used.

rate of movement / distance of fan from shoot

1/time in seconds / cm

0.016 200

0.018 175

0.023 150

0.034 125

0.055 100

0.071 75

0.092 50

0.11 25

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 Appendix 2

Practical 10 - Technical information

The effect of wind speed on transpiration in a leafy shoot

The apparatus and materials required for this practical are listed below.

The amount of apparatus listed is for one student or one group of students if they
are to work in groups.
1. 1 freshly cut leafy shoot that has been put immediately into fresh water
2. 1 potometer set up as shown in the diagram below.

3. electric fan
4. meter rule
5. sight of a stop watch or clock
Additionally each student will require access to a sink and running water.
Safety Precautions/Risks.
No specific hazards identified.

A risk assessment should be carried out as a matter of course.

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 Appendix 2

Practical 11 - Investigating the role of carbon dioxide in living organisms.

This practical focuses on manipulation and observations, recording and presenting

data, analysis, drawing conclusions and evaluating methods. The practical also
develops skills of using material in new and unfamiliar situations.

Intended learning outcomes

By the end of this practical you should be able to:

• Identify dependent and independent variables
• Make a hypothesis and express this in words
• Experience relevant methods, analysis and conclusion.
• Describe and explain the relationship between different living organisms and
the production of carbon dioxide
• Evaluate procedures

Safety Information

You should wear eye protection throughout this practical.

Ethanol is highly flammable. There should be no flames in the same room.

Bicarbonate indicator solution is flammable.

Background information
• Carbon dioxide is a gas found in the air at 0.04%
• Carbon dioxide dissolves in water to form carbonic acid thus reducing the
pH
• When bicarbonate indicator solution is equilibriated with air it turns
red/orange
• Bicarbonate indicator changes colour in different levels of pH
• You will remember from biology learnt in earlier courses that plants both
respire and photosynthesise.
• Respiration glucose + oxygen carbon dioxide + water + energy
• Photosynthesis carbon dioxide + water glucose + oxygen
• The point at which the carbon dioxide released by plants from respiration,
equals the carbon dioxide absorbed by plants for photosynthesis is called
the plant’s compensation point.

You will investigate the effect of different living organisms on bicarbonate indicator
and use this information to devise an experiment to determine the compensation
point in plants.

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 Appendix 2

• Read the information above

• Identify and write down the dependent and independent variables
• Write down what you think will happen (do not worry about what the colour
the indicator will be – you will discover that by doing the experiment)
• Identify any variables that should be controlled and outline how this should
be done
• Write down a hypothesis to explain what will happen to the colour of the
bicarbonate indicator when a plant is at its compensation point.

Method

Preparations and making observations

1. Rinse out three large test-tubes with distilled water and then with
bicarbonate indicator solution.
2. Using a syringe or small measuring cylinder place 3 – 5 cm3 of bicarbonate
indicator solution into each test-tube.
3. Carefully place a piece of perforated gauze in each test-tube so that it is
just above the indicator solution.
4. Place a rubber bung or cork into the first test-tube.
5. Carefully place three green seedlings onto the gauze in the second test-
tube and seal with a rubber bung or cork.
6. Carefully place three fly larvae onto the gauze in the third test-tube and
seal with a rubber bung or cork.
7. Place the three test-tubes near a bright light source such as a lamp or
window
green seedlings

larvae

gauze

bicarbonate indicator

8. Check that the colour of the bicarbonate indicator solution in each test-
tube is red/orange at the start of the experiment.
9. Leave the tubes for at least 30 minutes, comparing the colour of each
indicator solution every ten minutes.
10. When the colours look different in all three test-tubes, note the final colour
of the indicator in each of the three test-tubes.

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 Appendix 2

Write-up
• Record your results in a clear table.
• Explain why one of the test-tubes contained no living material
• Explain your findings using your knowledge of respiration and
photosynthesis
• Assess the reliability of the results obtained and suggest any modifications
you could make to improve the experiment
• Plan and describe, but do not carry out an experiment using the same
technique, to determine the compensation point in plants.

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 Appendix 2

Practical 11 - Lesson Plan

Investigating the role of carbon dioxide in living organisms.

Context

A practical investigation set in the context of 9700 syllabus – Gaseous exchange

Key aims of the lesson

This practical is designed to develop the skills of observation, analysis and evaluation
and using knowledge gained in a new and different context

Intended learning outcomes

By the end of the practical and the write-up the student should be able to
• Experience relevant methods, analysis, conclusions and evaluation.
• Describe and explain how an experimental method can be adapted to
discover when a plant is at its compensation point.

Resources required

White board or flipchart and suitable pens or blackboard and chalk

Practical materials specified on the Technical Information Sheet.

Copies of the student worksheets.

Planned activities

Timings/ Teacher/ Student Activities

minutes

End of Preparation – 2 page student worksheet given out for students to

previous read in preparation for the practical lesson. To consider identification
lesson of the variables, formulate a hypothesis and review previous learning
on cell membranes

0-3 Introduction to the aims, intended outcomes and shape of the lesson
– teacher led oral presentation

3-5 Context – review of pH indicators such as litmus and universal

indicator and that carbon dioxide dissolves to form an acidic solution.

5-8 Introduction to method – Teacher briefly outlines method and

answers any student questions on procedure. Teacher emphasises
safety concerns and ethics when handling living material such as fly
larvae which must not suffer undue stress.

8 - 40 Carrying out the practical – students carry out the practical work..

40 - 50 Obtain results – Students enter results into table and clear away
apparatus as soon as they have finished

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 Appendix 2

50 - 60 Drawing together the threads – Teacher led discussion on the skills

that have been developed as well as discussion on results obtained.
Practical write up to be completed in following lesson or as homework
activity

Useful information
Discussion / evaluation points should include:
• what colour bicarbonate indicator tuned in different situation
• the cause of the colour change in the bicarbonate indicator
• what other variables could have affect the results and which variables should
be controlled
• how the procedure could be improved to increase reliability
• how the procedure could be modified to determine the compensation point in
plants

A numerical value of the compensation point can be determined by using a light

metre. The light reading should be taken as close to the plant as possible at the time
when the plant is at its compensation point.
For students unable to obtain accurate data, the following table of results may be
used.
no living material seedlings fly larvae

colour of start red/orange red/orange red/orange

bicarbonate
indicator
end red/orange purple yellow

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 Appendix 2

Practical 11 - Technical information

Investigating the role of carbon dioxide in living organisms.

The apparatus and materials required for this practical are listed below.
The amount of apparatus listed is for one student or one group of students if they are
to work in groups.
1. 3 large test-tubes each fitted with a rubber bung or cork
2. gauze or similar to support the specimens in the test-tubes whilst at the
same time allowing the transfer of gases
3. supply of distilled water to rinse each test-tube.
4. 40 cm3 of bicarbonate indicator solution, sufficient to rinse each test-tube
and have sufficient remaining to place 5cm3 into each test-tube.
The stock solution of indicator can be prepared by dissolving 0.2g of thymol
blue and 0.1g of cresol red in 20cm3 of ethanol. Also prepare a solution by
adding 0.84g of pure sodium bicarbonate to 900 cm3 of distilled water. Add
the dyes to this solution and make up to 1 dm3. To prepare the indicator for
use, pipette 25cm3 of stock solution into a graduated flask and make up to
250 cm3 with distilled water.
The solution should be equilibriated with air by aspirating atmospheric air
through the solution until it is orange/red in colour.
5. 3 germinated seeds such that they have developed green leaves and are
photosynthesising. Cress seeds that have been placed on moist cotton wool
in a Petri dish will germinate and develop leaves in only a few days. Times
will vary depending upon local conditions.
6. 3 large fly larvae that are active and not approaching pupation
7. 10cm3 graduated pipette or measuring cylinder or syringe

Additionally each student will require access to a sink and running water.

Commercial bicarbonate indicator solution is available from most chemical

wholesalers, however it is possible to make up the solution in the laboratory as
described above.
Safety Precautions/Risks.

Ethanol = F

Bicarbonate indicator solution = F

A risk assessment should be carried out as a matter of course.

91
 Teaching AS Biology
Practical Skills

University of Cambridge International Examinations

1 Hills Road, Cambridge, CB1 2EU, United Kingdom
Tel: +44 1223 553554 Fax: +44 1223 553558 E-mail: [email protected] Website: www.cie.org.uk

© University of Cambridge International Examinations 2005

PSAS9700 0501