UNIVERSITY OF SANTO TOMAS

SENIOR HIGH SCHOOL

GENERAL BIOLOGY
1st Quarter, AY 2017-2018

LABORATORY EXERCISE # 3
Central Dogma of Molecular Biology

Background
Hereditary materials of most living organisms are contained in chromosomes which house long
fragments of deoxyribose nucleic acids, also called as DNA. The DNA double helix with its
sugar-phosphate backbone and nitrogenous bases is a highly organized material that can be multiplied and
transferred from parent to offspring through replication and transformed into proteins through translation
and transcription. The three processes mentioned summarizes the central dogma of molecular biology.

In this laboratory exercise, step by step process of DNA replication and protein synthesis through
RNA transcription and translation (initiation, elongation and termination) is further analyzed by
construction of a 3D model. Proper nucleotide base pairing will also be practiced with applications on
genetic mutation. Extraction of DNA from different fruits and inner cheek lining allows comparison of
DNA yield and actual observations of DNA fragments through the naked eye. Extensive application of
the central dogma in modern medicine and research will also be explored using an an online virtual
laboratory to extract DNA from Escherichia coli.

Materials

Fruits (enough for 1/2cup): Alcohol (Ethanol)

- Banana Barbecue stick
- Kiwi (10) Test tubes
- Strawberry (4) 100mL beakers
Knife (1) 500mL beaker
(3) Ziploc bags (1) Hot plate
Salt (3) Stirring rods
Strainer/cheesecloth (1) 10mL pipette
Tablespoon Plastic cups (one per student)
Liquid detergent Triple beam balance

Procedures

Extraction of DNA from a fruit

1. Peel the bananas and the kiwi. Remove the sepals from your strawberry. Cut into smaller pieces
and place 1/2 cup of each fruit in separate resealable plastic bags. Make sure to wash your plastic
knife before slicing different fruit.
2. Add a ½ spoon of salt and about 20 ml warm water (just enough to cover the fruit) to each bag.
Squash the fruit until there are no lumps.
3. Pour mixture in separate beaker through a strainer/paper towel/cloth. Make sure to use a clean
strainer/paper towel/cloth in every fruit.
 4. Add 1 spoon of liquid soap to each mixture and gently stir without creating bubbles.
5. Carefully add 20 ml of alcohol to each mixture. Wait for 5 minutes or until layers are formed.
6. With a stick, collect as many DNA fragments on top layer and transfer to separate tubes
containing 5 ml cold alcohol. Make sure to use different sticks for each fruit. Label each tube with
its corresponding fruit.
7. Compare the amount of DNA collected from each fruit.
8. Throw used disposable materials and clean your area.
9. Insert pictures and write observation on designated tables.

Extraction of DNA from Buccal Cells

1. Place 5 ml of 25% liquid detergent to the test tube (1 tube per student)
2. Gargle 10 ml of 0.9% salt water in your mouth for 30 seconds. Spit the water into your cup.
3. Pour your 10 ml collected buccal cell mixture to prepared tube. Slightly mix detergent and your
buccal cell mixture for 2-3 minutes without creating bubbles. Do not be too vigorous to avoid
breaking DNA into smaller fragments, a process known as shearing.
4. Open the tube and slightly tilt and pour 1 teaspoon (5 ml) of the chilled alcohol. Set for 5 minutes
or until it formed layers.
5. Collect the long DNA fragments by swirling the barbecue stick. This is called DNA spooling.
Don’t mix of the ethanol and soapy layers. If too much shearing has occurred, the DNA
fragments may be too short to wind up, and they may form clumps instead. You can try to scrape
these out with the stick.
6. Place the spooled DNA fragments to 2.5mL microcentrifuge tubes containing cold alcohol. DNA
will stay solid at this state.
7. Throw used disposable materials and clean your area.
8. Answer the questions on the worksheets

Data and Results

Extraction of DNA from fruits and buccal cells

Insert picture of the corresponding steps from your experiment. Write observations.

strawberry banana kiwi

Mashing of fruits

insert picture insert picture insert picture

Observations: Observations: Observations:

Addition of detergent

Page 2 of 4
 insert picture insert picture insert picture

Observations: Observations: Observations:

Addition of cold alcohol

insert picture insert picture insert picture

Observations: Observations: Observations:

Final output

insert picture insert picture insert picture

Observations: Observations: Observations:

insert picture insert picture

Collected buccal cells Addition of buffer

Observations: Observations:

insert picture of final output from each member and label collected DNA fragment

Final output
Observations:

Extraction of DNA from Escherichia coli

Enumerate at least three important steps with screenshot pictures in extraction of DNA from Escherichia
coli

Page 3 of 4
Guide Questions (For the Discussion):
1. What happens if basepair substitution occurs during DNA replication?
2. What is the term for series of multiplication of a mutated gene?
3. Give one example of a genetic disorder. Briefly discuss its causes, symptoms (w/ picture if
possible) and cure. Attach first page of scientific journal article that is relevant to this disease.
4. What is the purpose of the following steps/materials:
● Mashing of fruits
● Soap/detergent
● Warm water
● Salt/ Sodium chloride
● Chilled alcohol
5. Which of the fruits obtained highest yield of DNA? Why?
6. Some strains of E. coli cause bloody diarrhea, anemia and kidney failure that may lead to death.
What are further medical and research applications that may be done after the extraction of DNA
from E. coli?
7. Application of the central dogma in medicine and biotechnology has helped in many beneficial
ways. On the otherhand, this knowledge is also said to be a “double-edged sword”, why?

References:

Starr, C. & McMillan, B. (2016). Human biology, 11th ed. Boston,USA: Cengage.
Sweeney, D. (n.d.). DNA isolation from strawberries retrieved from
http://www.caseciw.org/first_light_case/horn/strawberries/strawbdnaproc.html
Coldspring harbor laboratory (n.d.) Laboratory DNA Extraction retrieved from
http://labcenter.dnalc.org/labs/dnaextraction/dnaextraction_d.html
Bailey, R. (n.d.) How to extract DNA from a banana retrieved from
http://biology.about.com/od/biologylabhowtos/ht/dnafromabanana.htm
University of California (n.d.) SEP lessons: See your DNA. Retrieved from
http://www.seplessons.org/files/See%20your%20DNA.pdf

Page 4 of 4