Nursing Biochemistry

Laboratory Report

Nucleic Acid
Activity No. 11

Cuenco, Lara Faith O.

Dela Torre, Sherieca L.
Kamdon, Mohammad Yusuf A.
Locker No.36; NurBio Lab C

1st Semester, SY 2019-20

Nursing Biochemistry

Laboratory Report

RATIONALE
Nucleic acids are vital for cell functioning, and therefore for life. There are two types of nucleic
acids, DNA and RNA. Together, they keep track of hereditary information in a cell so that the cell can
maintain itself, grow, create offspring and perform any specialized functions it's meant to do. Nucleic
acids thus control the information that makes every cell, and every organism, what it is.

Nucleic acids, which are composed of nucleotides, are very large and complex organic molecules
that contain the genetic code for that organism. Two closely related are needed to transmit the genetic
information from parent off-spring: DNA and RNA. They also share structural similarities. Both DNA
and RNA are polymers of individual nucleotides. Each nucleotide has three components, namely, five
carbon ring sugar, phosphate group and nitrogen bases. Both DNA and RNA have four nitrogen bases
available to construct nucleotides.

Nucleic acids allow organisms to transfer genetic information from one generation to next. When
a cell divides, its DNA is copied and passed from one cell generation to the next generation. DNA is
organized into chromosomes and found within the nucleus of the cell. It contains the “pragmatic
instructions” for cellular activities. RNA is essential to the synthesis of proteins. It is also a component of
cell organelles called ribosomes. Information contained within the genetic code is typically passed from
DNA to RNA to the resulting proteins.

One of the many advantages to using yeast as model system is that large quantities of
biomolecules, including nucleic acids which are DNA (Deoxyribonucleic acid) and RNA (Ribonucleic
acid) can be purified from cultured cells (Isolating Nucleic Acids from Yeast. JoVE, Cambridge, MA.
2019).

Objectives:

• To isolate RNA from the yeast

• To identify the products of hydrolysis of RNA
• To determine the physical and chemical properties of the isolated yeast RNA

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry

Laboratory Report

DATA INTERPRETATION

Table No. 1

Name of Test Reagent Result

1. Nucleoproteins 10%NaOH, 1% CuS04 Blue to violet color

Theoretical Result: Violet color

with blue precipitate

Discussion

Biuret test is done to show the presence of peptide bonds, which are the basis for formation of
proteins. These bonds will make the blue biuret reagent turn to violet or purple color. Formation of the
purple color is due to reduction if Cu2+ to Cu+. (BiologyNotes. 2019).

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry

Laboratory Report

Table No.2

Name of Test Reagent Result

2. Mild Acid Hydrolysis 10% H2SO4 Light brown/Champagne color

a.Presence of Phosphate 10% HNO3, 5% ammonium Turned Yellow color

Molybdate
Theoretical Result: Yellow color
with Yellow precipitate

b.Presence of Ribose Bial’s Orcinol Reagent

#1 (Soln fr Acid Hydrolysis) #1 White Precipitate (Blue-green

condensation product)
#2 (0.1% Ribose)
#2 Cloudy precipitate
#3 (0.1% Glucose)
(Blue green-condensation
product)

#3 Brown Precipitate

c.Presence of Purines 10%NH4OH, 5%AgNO3 The solution turned pink color

and formation of white
precipitate

Theoretical result: Foamy

gelatinous substance precipitate

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry

Laboratory Report

Discussion

Mild Acid Hydrolysis:

Presence of Phosphate

The test for the presence of phosphate is the test for inorganic phosphate or ammonium
molybdate test hydrolyze the pyrophosphate to phosphate and react with the reagent added and produces
yellow color and yellow precipitate (Gosiengfiao, J. 2019).

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry

Laboratory Report

Presence of Ribose

Bial’s test is a test for the presence of pentose. It can be used as a test for RNA due to the
presence of ribose. Bial’s reagent consists of reagents which promotes the dehydration of ribose to
furfural. The presence of furfural can be detected by the addition of a test reagent, known as the
condensation reagent. The condensation reagent is generally a phenolic compound that reacts with
furfural to give a highly-colored product. In the experiment, Sodium hydroxide and orcinol as the
condensation reagent. A positive test for pentose is indicated by a blue-green condensation product (Rao,
1992). For glucose, it gave a brown color solution indicating the absence of ribose in the solution (Course
Hero. n.d.)

Presence of Purines

Hydrolysis of N-β-glucosidic bonds between purine bases and ribose or deoxyribose results in a
release of purine bases (A&G) caused by NH4Oh. Ag+ precipitate cause the formation of foamy
gelatinous substance precipitate (Gosiengfiao, J. 2019).

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry

Laboratory Report

ANSWERS TO QUESTIONS

1. Name the purine bases found in nucleic acid

Nucleic acids consist of chains of linked units called nucleotides. In detail, the four
nitrogenous bases that form the nucleotide units of a DNA strand are two purines (adenine [A]
and guanine [G]) and two pyrimidines (cytosine [C] and thymine [T]).

2. Account for the information of the precipitate in the test for purines
The role of silver nitrate/basic condition is to precipitate purines via its interaction with
the purines nitrogens. Test for purines: Add excess 2M ammonia solution and a few drops of
0.1M silver nitrate to 1 mL of DNA extract. A white precipitate indicates the presence of purines.
The precipitation mechanism is by a reaction of Ag+ with the nitrogens of purines.

3. What is the basis of Bial’s Orcinol test?

Bial's test is a chemical test for the presence of pentoses. It is named after Manfred Bial, a
German physician. The components include orcinol, hydrochloric acid, and ferric chloride. A
pentose, if present, will be dehydrated to form furfural which then reacts with the orcinol to
generate a colored substance.

4. What is ATP? Why is it important in the cell?

ATP – or Adenosine Triphosphate – is the primary energy carrier in all living organisms
on earth. Microorganisms capture and store energy metabolized from food and light sources in
the form of ATP.
When the cell requires energy, ATP is broken down through hydrolysis. The high energy
bond is broken and a phosphoryl group is removed. The energy released from this process is used
to drive various cellular processes. ATP is constantly formed and broken down as
it participates in biological reactions and it is central to the health and growth of all life. Without
it, cells could not transfer energy from one location to another, making it impossible for
organisms to grow and reproduce!
Since ATP is present in all living and active microbial cells, it is an excellent indicator of
overall microbiological content in fluids or deposits. By measuring ATP regularly, and being able
to differentiate between cellular ATP inside active microorganisms and dissolved ATP released
from dead cells, cause & effect relationships can be identified helping you solve microbiological
challenges before it’s too late.

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry

Laboratory Report

CONCLUSION

Isolation of RNA from yeast involves heating with NaOH because it loosen and lysed the cell
membrane resulting in the extraction of RNA. Addition of NaOH and glacial acetic acid to the sample of
RNA prevents its degradation by increasing its pH level which inactivates the nucleases (Guse, I. 2014).

RNA can be extracted from the yeast by rupturing the yeast cells, increasing the pH level to
denature unwanted and contaminant proteins and substances and centrifugation to separate the denatured
substances and the unwanted substances. There are also different tests to determine the content and
composition of nucleic acids in this case, the RNA. In the experiment, the hydrolysed RNA is positive for
phosphate, ribose and purine bases.

Characterization test are used to describe the reactions of RNA to different reagents in order to
represent the structural features. According to research, for the ribose test, the orcin reaction is a test for
the presence of ribose in RNA. The reaction depends on the conversion of ribose to an aromatic aldehyde
(furfural) which then reacts with the orcin to give blue-green color. For the test for the presence of
phosphate in the hydrolysed RNA, the positive result is a yellow precipitate. The positive result is
because of the reactuion of the ammonium molbdate solution, which when dropped onto the sample will
result to a yellow stain which indicates the presence of phosphate or a crust of phospo-ammonium
molybdate. The test for purines gives the reaction of RNA with ammonium hydroxide given that purines
are known to be readily soluble in dilute acids. Yellow precipitate upon evaporation resulted from the
oxidation process.

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry

Laboratory Report

REFERENCES

Shafer, J. (2017, October 25). What is the importance of nucleic acid?. Retrieved from:
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Blanco, A. (2017). Nucleic Acids. Retrieved from:

https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/nitrogenous-base

Karaman, R. (2016, October 10). Test for purines. Retrieved from:

https://www.researchgate.net/post/What_is_the_role_of_ammonium_hydroxide_and_Silver_nitrate_in_th
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Donaher, A. (2016, October 19). What is ATP and what does It do? Retreieved from:
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Why is ATP important? (n.d.). Quizlet. Retrieved from: https://quizlet.com/66792002/why-is-atp-

important-flash-cards/

Biuret test for proteins. (n.d.). Weebly. Retrieved from: https://biology-igcse.weebly.com/food-

test-4---biuret-test-for-proteins.html

Gosiengfiao, J. (2017, June 22). Nucleic Acids. Retreived from:

https://www.scribd.com/document/351991577/Exp-10-Biochem

Isolating nucleic acids from yeast. (2019). Jove Science Education Database. Retrieved from:
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San Juan, G. (2013, February 10). Isolation of RNA from yeast. Retrieved from:
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Janina, M. (2011, March 7). Nucleic Acids. Retrieved from:

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Gilig, Lloyd. (2017, January 13). Nucleic Acids. Retrieved from:

https://www.scribd.com/document/338035486/Experiment-3-Nucleic-Acids

Balda, K. (2011, September 11). Nucleic acid, RNA. Retrieved from:

https://www.slideshare.net/kevbalda/report-exp-6-and-7-dna-and-rna

Santos, R. (n.d.). Bial’s test for the presence of pentose. Retrieved from:
https://www.coursehero.com/file/p8v62j/Bials-test-is-a-test-for-the-presence-of-pentose-It-can-be-used-
as-a-test-for/?fbclid=IwAR3eqAgVX-zVFXq6aNfV4isdaKTD3MnH8qXquEc5OG1ol9i1m-
0qIWXxHdM

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University
Nursing Biochemistry

Laboratory Report

Poblete, C. (2013, September 3). Nucleic Acid. Retrieved from:

https://prezi.com/fzxbr2rtl_xb/experiment-10-nucleic-acids/?fbclid=IwAR1G-
ObDXf_TaUxGQf3bpLvg9w5WdxGQiCT4oZyNv3IAPgVWUywggX2D-Bs

Sundin (n.d.). Isolation of Casein: Protein and Amino Acid. Retrieved from:
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Acid?fbclid=IwAR2ubyGFUX3jrqp2-nq0KdeeizIpE1fUWMwTrhhB4VMNmoZmy_DkDGifAjA

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Natural Sciences Department, College of Science and Information Technology, Ateneo de Zamboanga University