Lab Report in Nucleoproteins

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Polytechnic University of the Philippines

College of Social Sciences and Development


Department of Psychology
Nucleoproteins
Group 1, BS Psyhology 3-3
Acosta, Sheene Khristia, Agdeppa, Tiffany Mae,
Alipao, Ma Erika, Aniban, George Jr., C.,
Ayoso, Trisha, Banco, Nerissa, Barrento, Allyza Khate

Abstract
In this experiment, nucleoproteins that were collected from the filtrate of banana
smoothie that was placed in a test tube with ice cold ethanol were used to analyze the purity of
the DNA sample of a banana. A smoothie composed of banana, ice, and saturated sodium
chloride solution was filtered with a cheese cloth and 80 ml of the filtrate was equally distributed
to 8 test tubes and ice cold ethanol was poured after, then the tubes were planted in a container
filled with crushed ice so that the nucleoproteins would float to the surface and it would be easier
to collect. The purity of the DNA was measured by using a Spec photometer, the purity of the
isolated DNA sample that was tested is not a good quality because the ration of purity is only at
1.8-2.0 and the result of the experiment is 1.108. The experiment was considered to be a success
even though it yielded a not so high ratio of purity because they were still able to get
nucleoproteins from the samples so the technique that was used is still effective.

I. Introduction
Nucleic acids are molecules that structure of the purine and pyrimidine bases.
allow organisms to transfer genetic UV radiation exposure can cause DNA
information from one generation to the next. damage and mutations. However these
There are two types of nucleic acids they are absorption properties of DNA can be used
deoxyribonucleic acid DNA and ribonucleic for detection, quantification, and assessment
acid RNA. Simple protein bonded to it is of purity. Pure DNA preparations have an
called nucleoproteins. Proteins inside the A260/ A280 ratio greater than or equal to
cell have many functions such as building 1.8.
cellular structures and catalysts for cellular
reactions that give cell its specific Studying nucleic acid or molecules
characteristics. that code life are the primary work of
genetic engineers. These scientists involved
Why DNA absorbs UV? The reason in this program follow s careful protocols to
that DNA absorbs UV radiation is that produce valuable medicines. The results of
energy of the UV photon matches the understanding the biochemistry of nucleic
difference between two different energy acis create products which can be use for
states of DNA’s electronic structure. DNA is betterment of human, but we should be
very complex molecule electronically; the careful because a few differences in coding
part of the molecule that is thought to absorb of these molecules can create great change
most of the UV radiation is the aromatic ring in an organism.
II. Methodology and Materials III. Results and Discussion
The materials that were used are Nucleoprotein, conjugated protein
eight (8) test tubes, two (2) cubets, beaker, consisting of a protein linked to a nucleic
wire loop, petri dish (we used petri dish acid, either DNA (deoxyribonucleic acid) or
instead of a watch glass), test tube rack, and RNA (ribonucleic acid). The protein
a blender. The materials that we brought are combined with DNA is commonly either
distilled water, 95% ice cold ethanol, ice, histone or protamine; the resulting
dishwashing liquid, banana and a cheese nucleoproteins are found in chromosomes.
cloth.
On this experiment, we smashed the
The first step in the experiment was banana with distilled water into a blender.
blending the banana with ice to make a fruit Then we added 10 mL of dishwashing liquid
shake. After the blending process, 200 mL and 200 mL of saturated sodium chloride
were measured and placed in a beaker. This solution.
will become the sample in DNA isolation.
The next was placing back the 200 mL
sample in the blender while adding
approximately 10 mL dishwashing liquid
and 200 mL saturated sodium chloride
solution. The saturated sodium chloride
solution was created by mixing 15 g of rock
salt and 200 mL of distilled water. When the
mixture inside the blender is properly mixed,
it was filtrated using a cheese cloth in a
beaker. After achieving about 150 mL of
filtrate, it was put into 8 test tubes,
approximately 10 mL for each test tube. It
was overlayed with 8 mL of ice cold Once the dish soap helps release the
alcohol. All of the test tubes were put in a DNA, this salt will help the DNA strands to
cooler with ice to separate the stick to each other in clumps large enough
nucleoproteins. for us to see. Dish soap can break apart a
type of molecule called lipids. Think of fats
After waiting for the nucleoproteins
and oils. Dish soap breaks down those
to float in the alcohol, it was spooled using a
greasy molecules. Now, the molecules that
wire loop and was placed in a petri dish. The
make the membranes around cells and the
nucleoproteins that are collected were then
nucleus (which holds DNA) are lipids. So
mixed with 10 mL distilled water. The next
when dish soap is added, the cell membrane
step was putting the distilled water with
and the nuclei are broken apart, releasing the
nucleoproteins in one of the cubets. The
DNA.
other cubet was placed with distilled water.
Both of the cubets were then placed in a After that, we filtered the mixture
spectrophotometer to measure its absorbance using the cheese cloth and then place the
at A260 and A280. mixture in 8 test tubes. Then, we put a cold
ethanol filling up 8 mL to the mixture. The
DNA clumps are soluble in some liquids,
but not in alcohol. So adding ethanol helps
the clumps of DNA to form. The cheese
cloth serves as the “filter” in the experiment. means that it separates from other materials
The layers of cheesecloth trap the in the liquid. The salt was needed in the
precipitated cell debris of the banana while procedure because together with the
the soluble DNA passes through. dishwashing soap, each worked to dissolve
the bilipid layer of membranes and release
the cellular contents, including DNA.

After the filtration process, the


products formed were a purer sample of
DNA extract. On the other hand, by adding
iced cold ethanol to the DNA filtrate The purine and pyrimidine bases in
solution in the tube, the DNA is rendered DNA strongly absorb ultraviolet light.
visible. Ethanol dehydrates the DNA by Double-stranded DNA absorbs less strongly
removing the water. This dehydrated than denatured DNA due to the stacking
molecule then forms a precipitate, which interactions between the bases.
It is important to measure the DNA nucleoproteins in a cold ethanol is that DNA
sample with A280 and A260 so that we is insoluble in alcohol, so using ethanol
cannot just only base in the purity of the causes the DNA to precipitate out. The
DNA alone, but also the preciseness of our colder the ethanol, the less soluble the DNA
nucleic acid isolation. Based on our results, is.The colder the ethanol is the greater the
the purity of our isolated DNA is not in a amount of DNA that is precipitated. We
good quality since the ration of purity is conclude that we were able to conduct the
only at 1.8-2.0 and the result we got is experiment about nucleic acids through
1.108. Isolation of DNA from a Fruit Smoothie
successfully.
The purpose of molecule guanidium
chloride is to help destroy the tertiary
structure of the proteins that can be found in
the DNA solution. V. Recommendation
For the experiment to be effective,
the bananas that were supposed to use must
IV. Conclusion be Lacatan. It is hard to get proteins from
other kinds of banana such as Senorita. It is
We conducted an experiment to learn important to mix the ingredients well. While
more about nucleic acids through the filtering the filtrate, it is easier to mix it
process of isolation of DNA from banana continuously. The mixture must be always
smoothie. We gathered nucleoproteins by mixed, so that the bubbles will mix to the
collecting filtrate from the said smoothie mixture and not be stuck up on the surface.
composed of banana, ice, dishwashing liquid Make sure that instructions will be done
and alcohol using cheese cloth. We properly. Make sure that the measurements
transferred the collected filtrate in a test tube are correct and the way how it should
and poured ice cold ethanol on it. We measure must be double checked. One
planted the test tubes afterwards in a mistake in instructions might cause changes
container full of crushed ice to maintain the in results too.
temperature. We collected the floating
nucleoproteins using wire loop and
transferred it to petri dish then to test tube
with distilled water. We also tested the
absorbance at A260 and A280 using
spectophotometer. Spectrophotometry is a
method to measure how much a chemical References
substance absorbs light by measuring the http://www.mgp.cz/files/nanodrop/manualy/
intensity of light as a beam of light passes pomer_cistoty.pdf
through sample solution. We have the result
of:
A260 = 0.821
A280 = 0.741
A260/A280 = 0.821/0.741 = 1.108.
The reason why we were able to collect

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