Nursing Biochemistry

Laboratory Report

Nucleic Acids
Activity No. 11

Patega, Jade K. (Principal Author)

Macion, Kristine
Sumido, Allen
Locker No.6; NurBio Lab A
2nd Semester, SY 2019-20
Nursing Biochemistry
Laboratory Report

RATIONALE

Nucleic acids are the most important biological macromolecules and are responsible for
encoding, transmitting and expressing genetic information. These molecules carry information
that enables the production of proteins and replication of the genetic material for the new cells.
Nucleic acids are large molecules loaded with small and fine details, the genetic data. Nucleic
acids occur in animals, plants, fungi, bacteria, viruses and any other living organism that uses
and converts energy. By comprehending the nucleic acids and its mechanism, we can
potentially understand how diseases happen and eventually its cure.
Nucleic acids are tiny molecules but they play a major role in the biology of the human
body. Scientists have discovered that diseases occur due to error in the genes and proteins, but
further understanding can help scientists decode how exactly diseases occur and ultimately how
to cure them (Seeley, 2015.)

The objectives of this experiment are 1.) to be able to isolate RNA from yeast, 2.)to determine the
presence of the following: phosphates, ribose and purines and 3.) to determine the reduction
ability of sugar.

DATA INTERPRETATION

Table No.1 Test for Nucleo-Proteins

Test Solutions Added reagent Observation

+ 10% NaOH Violet with blue precipitate

Filtrate ( Yeast-RNA)
+ 1% CuSO4 (Positive)

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

Biuret’s Test is a chemical test used for detecting the presence of peptide bonds. In a
positive test, a copper (II) ion is reduced to copper (I), which forms a compound with the
nitrogens and carbons of the peptide bonds in an alkaline solution. A violet color means positive
and indicates the presence of proteins while, light blue indicates that no peptide bond is present.

Table No. 2 Test for Phosphates

Test Solutions Added Reagent Observation

+ 10% HNO3 Yellow – green precipitate

Acid Hydrolysate
+ 5% Ammonium (Positive)

Ammonium molybdate is a white, crystalline salt used as an analytic reagent, that acts as a
precipitant of phosphoric acid, and in pigments. It is used to detect phospates in nucleic acid. It indicates
the presence of phosphorus by a yellow stain when dropped upon a specimen. The yellow precipitate is
caused by the hydrolysis of pyrophosphate to phosphate.

Table No. 3 Test for Ribose

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

Test Solutions Added Reagent Observation

Acid Hydrolysate + Bial’s Orcinol Reagent Green (Positive)

+ Bial’s Orcinol Reagent Green (Positive)

0.1% Ribose

0.1% Glucose + Bial’s Orcinol Reagent Brown (Positive)

Bial’s test, is a chemical test used to detect the presence of sugars and their derivatives which forms
furfural upon heating in acidic medium. Furfural formed from pentoses reacts with orcinol in the presence
of FeCl3 to give off blue- green hue of the solution. Since ribose form furfural with a strong acid, it is
responsible for the positive result of RNA for this test. Both green and brown color indicates a positive
result. Furthermore, green colored product indicates a positive test for the presence of pentoses, while
brown colored product indicates the presence of hexoses.

Table No. 4 Purines Bases

Test Solutions Added Reagent Observation

+ 10% NH4OH White precipitate present

Acid Hydrolysate
+ 5% AgNO3 (Positive)

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

Silver nitrate helps precipitate purines through its interaction with the purine
nitrogen.10% NH4 and 5% AgNO3 are chemicals used to detect the presence of purines
through precipitation of Ag+ ions. The precipitation reaction is caused by the reaction of
Ag+ with the nitrogens of purines. In addition, hydrolysis of N beta glycosidic bonds
between purine bases both in ribose and deoxyribose results in a release of purine bases
such as adenine and guanine caused by NH4OH. White precipitate indicates a positive
result for the presence of purines. In addition, Ag+ precipitate causes the formation of
foamy gelatinous substance.

ANSWERS TO QUESTIONS:

1. Name the purine bases found in nucleic acids

A set of five nitrogenous bases is used in the construction of nucleotides, which in turn build
up nucleic acids like DNA and RNA. These nitrogenous bases are adenine (A), uracil (U), guanine
(G), thymine (T), and cytosine (C).
2. Account for the formation of precipitate in the test for purines.
The precipitation reaction is caused by the reaction of Ag+ with the nitrogens of purines. Silver
nitrate is also responsible for the formation of gellatinous subtance.
3. What is the basis of Bial’s Orcinol test? The test reagent dehydrates pentoses to form furfural.
Furfural further reacts with orcinol and the iron ion present in the test reagent to produce a bluish
product. A positive test is indicated by the formation of a bluish product. All other colors indicate
a negative result for pentoses. Moreover, hexoses generally react to form green, red, or brown
products.

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

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

Adenosine Triphosphate (ATP), or Adenosine-5'-triphosphate, is another important nucleic acid. ATP is

described as the "energycurrency" of the cell or the "molecular unit of currency." One molecule of ATP
contains three phosphate groups, and it is produced by ATP synthase from inorganic phosphate and
adenosine diphosphate (ADP) or adenosine monophosphate (AMP). The structure of ATP consists of the
purine base adenine, attached to the 1' carbon atom of the pentose sugar ribose. Three phosphate groups
are attached at the 5' carbon atom of the pentose sugar. It is the removal of these phosphate groups that
convert ATP to ADP (adenosine diphosphate) and to AMP (adenosine monophosphate).
5. Show the structures of AMP, ADP and ATP.

AMP ADP

ATP

CONCLUSION:

RNA can be extracted from yeast by rupturing the yeast cells, increasing the pH is
done in order to free them from unwanted proteins that may disturb the desired
result and centrifugation to separate the desired from unwanted products. Several
test can be done to determine the content and composition of nucleic acids,
specifically the RNA. In the following test that was done, the hydrolyzed RNA is
positive for Biuret’s test, Bial’s test, Phosphate test and Purines test. The desired
objectives for the activity were met.

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

REFERENCES
 Drug discovery in nucleic acid、Nucleic acid medicine - BONAC corporation. (n.d.).

Retrieved from http://www.bonac.com/global/en/nucleic/about/

 Helmenstine, A. M. (2019, May 9). What ATP Is and Why It's Important in Metabolism.

Retrieved from https://www.thoughtco.com/atp-important-molecule-in-metabolism-

4050962

 Arvinluissantos. (n.d.). Bials test is a test for the presence of pentose It can be used as a

test for: Course Hero. 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/

 Nitrogenous Base. (n.d.). Retrieved from

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

biology/nitrogenous-base

 Nucleic Acids. (n.d.). Retrieved from

https://www.sciencedirect.com/topics/neuroscience/nucleic-acids

 Shafer, J. (2019, March 2). What Is the Importance of Nucleic Acids? Retrieved from

https://sciencing.com/what-is-the-importance-of-nucleic-acids-12186031.html

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