EXPERIMENT NO.

9
NUCLEIC ACIDS
INTRODUCTION
Nucleic acids are found in all living cells. It plays a
vital role in cellular organization and function. It plays an
integral role in cell division, reproduction, and
transmission of hereditary molecule functions that
functions as co-factors and co-enzymes.

Nucleic acids are mildly soluble in cold water,

insoluble in alcohol, but mixes instantly in weak alkali
forming alkali metal salts. They precipitate from the
alkaline solution by the addition of acid.
INTRODUCTION cont’n
In isolating RNA from yeast, heating with alkali is
essential. This extracts the nucleic acids and water-soluble
proteins and inactivates the nucleases. The nucleic acid is
separated from associated protein and other interfering
substance by acid extraction at pH 4.5. The final step is
treatment with alcohol with concentrated HCl to
precipitate the RNA followed by repeated washing with
alcohol and other substances that may interfere with the
chemical tests.
OBJECTIVES

1. To isolate RNA from yeast.

2. To test the properties of the isolated RNA.
MATERIALS
• Mortar and pestle • Bunsen burner
• 4 beakers (250 mL) • 8 test tubes
• Graduated cylinder • Test tube rack
• Pipette • Cheese cloth
• Aspirator • Filter paper
• Watch glass • 5% ammonium molybdate
• Tripod • 0.1% ribose solution
MATERIALS
• 0.1% glucose • 0.2% NaOH
• Bial orcinol reagent • 10% NaOH
• 10% NH4OH • 1% CuS04
• 5% AgNO3 • 10% H2S04
• Yeast • 10% HNO3
• White sand
PROCEDURE
A. ISOLATION OF RNA FROM YEAST
Mix and grind 2 grams of yeast with 2 grams of white
sand in a mortar. Then add 15 mL of freshly prepared 0.2%
NaOH to make a smooth creamy paste. Pour the mixture in
a 250 mL beaker and dilute with 0.2% NaOH solution to
make 50 mL. Cover the beaker with a watch glass to avoid
evaporation. Heat the beaker in a water bath with a
constant temperature of 90°C for 30 minutes. Filter the
solution thrice using cheese cloth and once through filter
paper. Allow the filtrate cool. Perform the following test on
the filtrate.
B. QUALITATIVE TEST

1. Test for Nucleoprotein

Place 1 mL of filtrate in a test tube, then add 1 mL of
10% NaOH solution and 5-10 drops of 1% CuSO4
solution. Notice the color produced.
OBSERVATION
ANALYSIS
B. QUALITATIVE TEST

2. Mild acid hydrolysis

Place 20 mL of 10% H2SO4 to the remaining filtrate

in a beaker. Boil the solution gently for a few minutes.
Perform the following test on the solution.
Continuation

a) Test for Inorganic Phosphates

Add 1-2 mL ammonia to 1 mL of the filtrate. Acidify
it using HNO3 then add 2 mL of ammonium molybdate.
Boil the solution then allow it to stand for a few minutes.
Note the color of the precipitate.
OBSERVATION
ANALYSIS
CONTINUATION
b) Test for the presence of ribose (pentose)

Prepare 3 test tubes and place the following

solutions as indicated below.
Test tube no. 1: 1 mL 0.1% ribose solution
Test tube no. 2: 1 mL 0.1% glucose solution
Test tube no. 3: 1 mL solution from the acid
hydrolysis
CONTINUATION

Add 3 mL of Bial orcinol reagent to each test tube.

Place the test tubes in a boiling water bath until the color
changes. Compare the color formed with the one that
contains the acid hydrolysis.
OBSERVATION
ANALYSIS
CONTINUATION
c) Test for the presence of purines.

Add 3 mL 10% NH4OH to 2 mL of the filtrate in a test

tube. Mix 2-3 drops of 5% AgNO3 solution to it. Note the
color of the precipitate formed.
OBSERVATION
ANALYSIS
ANALYSIS
 TESTS OBSERVATION EXPLANATION
Nucleoproteins (filtrate
from yeast RNA)
Purine Bases (acid
hydrolysate)
Test for ribose

Acid hydrolysate

0.1% ribose

0.1 glucose

Phosphates (acid
hydrolysate)
QUESTIONS
1. Why is yeast used as a source of RNA?

2. Name the purine bases found in nucleic acids?

3. Account for the formation of precipitates in the test

tube for purines?
CONCLUSION