BIOCHEMISTRY

NUCLEIC ACID
Presented by: Group 2
Nucleic Acids
• Nucleic acids are chemical molecules that are made up
of nucleotides. Each nucleotide is made up of three
parts: a pentose sugar, a nitrogen base, and phosphate.
There are two types of nucleic acids based on the
pentose sugar present: DNA with deoxyribose sugar
and RNA with ribose sugar. The presence of
phosphorus as phosphate, pentose sugar, and nitrogen,
which are purine and pyrimidine derivatives, is used to
conduct qualitative studies of nucleic acids.
Isolation of RNA from Yeast
• Fill a beaker with 125ml of water. Mix with 25 mL of
1% NaOH and 20 g of dry yeast. Heat the ingredients
in a water bath for 30 minutes, stirring regularly. Take
the mixture out of the water bath and strain it through a
cheese cloth. Allow the filtrate to cool before adding 3
drops of aceticacid. Evaporate the filtrate until it
reaches a volume of 50 mL. To precipitate the RNA,
cool and vigorously mix in 100 ml of 95 percent
ethanol with 1 ml of strong HCI. When the RNA has
settled, decant it and dry it in a petri plate at room
temperature.
SOLUBILITY TEST
REAGENTS: ALCOHOL, DILUTE HCI , DILUTE NAOH

1. Place in a separate test tube 1 ml each of the

following: cold water, hot water, alcohol, dilute HCI, and
dilute NaOH.

2. Add a pinch of the yeast RNA to each test tube. Shake

well. Observe.
Result:
- Cold water - insoluble
- Hot water - slightly soluble
- Alcohol = slightly soluble Dilute HCI - insoluble
- Dilute NaOH - very soluble
HYDROLYSIS OF RNA IN ACID
REAGENTS: 10% H2SO4
• PROCEDURE:
1 . Place a small amount of the yeast RNA in a test tube.

2 . Add 10 ml of 10% H2SO4, cover, and boil in a water

bath for 30 minutes.

3 . Divide the contents of the test tube into 4 equal

portions. Label the test tubes 1, 2, 3 and 4. Perform the
following tests below.
BENEDICT’S TEST
R EAG E NT S : S O LI D NA2 C O 3 , B E N E D I CT ’S S O L
UTION
• Neutralize the solution in test tube 1 with solid Na2CO3, let
stand for 2 minutes and then decant.

• Add 1 ml of Benedict’s solution in the test tube and place in a

water bath for 5 minutes. Observe .
Results:
Positive Benedict’s Test:
Formation of a reddish precipitate within three minutes. Reducing
sugars present.

Example: Glucose

Negative Benedict’s Test:

No color change (Remains Blue). Reducing sugars absent.

Example: Sucrose
TEST FOR PURINE REAGENTS:
NH4OH 1 %, AGNO3

1. Treat the solution in test tube 2 with 3 drops of ammonia water

to make it alkaline.

2 . Add five drops of 1 % AgNO3 solution and let stand for five
minutes. Observe.
White precipitate - positive
Reagents: Bial' s Reagent
TEST FOR PENTOSE REAGENT: BIAL’S
REAGENT.

PROCEDURE :
1.Add the solution in test tube 3, add 5 ml of Bial’s reagent.

2.Heat test tube in a boiling water bath for 10 minutes. Observe.

Results:
Positive test is indicated by the formation of a bluish
product.
TEST FOR PHOSPHATE REAGENT:
AMMONIA WATER, 6NHNO3, ( NH4 ) 2MOO4 SOLUTION

1.In test tube 4, and add five (5) drops of ammonia water,
acidify with 2 drops of 6N HNO3.

2.Add 1 ml of (NH4)2MoO4 solution.

3.Heat the mixture in a water bath for 5 minutes.

Observe.
Result:
The result of the test for phosphate is green on top,
then blue on bottom and also in unhydrolyzed almost
the same result.
REPEAT THESE TESTS USING UNHYDROLYZED RNA
AND COMPARE THE RESULT BY COMPLETING THE
TABLE:

HYDROLYZED RNA UNHYDROLYZED RNA

BENEDICT’S TEST Black/yellow Cloudy

TEST FOR PURINE White Cloudy

TEST FOR PENTOSE Brown/Furfural(Colorless) Cloudy

TEST FOR PHOSPHATE green/blue/Yellow Cloudy

Thank You!