TEST FOR NUCLEIC ACIDS &

ITS COMPONENTS

GROUP 4 – SECTION B

HO – USERO – UY – VALDEZ – VALENCIA – VARGAS – VILLARIN –YATCO – ZARATE

 NUCLEIC ACIDS

• Informational molecule
• Two kinds:
• DNA – deoxyribonucleic acid
• RNA – ribonucleic acid
• DNA – blueprint of living organisms
• RNA – machinery for the expression of the coded information
in DNA to proteins
• mRNA – carrier of genetic information encoded in the DNA

EXPERIMENT 6: TEST FOR NUCLEIC ACIDS AND ITS COMPONENTS

 KINDS OF NUCLEIC ACIDS

1. DNA
2. RNA
• Primary function of DNA – storage of genetic material
• Central Dogma of LIFE: flow of genetic material
• DNA → mRNA → protein
↑
Replication → transcription → translation

EXPERIMENT 6: TEST FOR NUCLEIC ACIDS AND ITS COMPONENTS

 COVALENT STRUCTURE OF
NUCLEIC ACIDS

• Genetic information is encoded in DNA and carried by mRNA as sequence or

a chain of nucleotides
• Composed of polymers of nucleoside monophosphates (nucleotide-
nucleotide-nucleotide)
1. Monomeric units consist of: NUCLEOTIDE
Base-sugar-phosphate (BSP)
a. Nitrogenous base which may be a purine or a pyrimidine
b. A pentose sugar, ribose or deoxyribose, in a furanose ring form
c. A phosphate group esterified to the sugar

EXPERIMENT 6: TEST FOR NUCLEIC ACIDS AND ITS COMPONENTS

 COVALENT STRUCTURE OF
NUCLEIC ACIDS

• Nucleoside-coupling of a base and a sugar

• Nucleotide – when a nucleoside becomes phosphorylated
• Nitrogenous base
• Purines – Adenine (A), Guanine (G)
• Pyrimidines – Cytosine (C), Uracil (U), Thymine (T)
• Sugar
• DNA – Deoxyribose
• RNA – Ribose
• Sugar hold the base on one side and the phosphate on the other side
• Thus, sugar hold the components of the nucleotide together.

EXPERIMENT 6: TEST FOR NUCLEIC ACIDS AND ITS COMPONENTS

 COVALENT STRUCTURE OF A
NUCLEOTIDE

EXPERIMENT 6: TEST FOR NUCLEIC ACIDS AND ITS COMPONENTS

 A. ISOLATION OF RNA FROM YEAST

IN A 250 mL
MIX 2 GRAMS OF ADD 15mL OF 0.2% BEAKER ADD
GRIND THE
YEAST WITH 2 NaOH TO MAKE A ENOUGH 0.2%
MIXTURE IN A
GRAMS OF WHITE SMOOTH CREAMY NaOH SOLUTION
MORTAR
SAND PASTE TO MAKE THE
VOLUME 50mL

COVER THE BEAKER FILTER 3 TIMES

WITH WATCH GLASS. THROUGH CHEESE COOL THE
HEAT IN A WATER CLOTH AND ONCE
BATH CONTROLLED
FILTRATE.
THROUGH FILTER
AT 90 C FOR 30 MINS. PAPER
 1. TEST FOR NUCLEOPROTEINS

1 mL OF ADD 1 mL OF ADD 5-10 NOTE THE

FILTRATE IN A 10% NaOH DROPS OF 1% COLOR
TEST TUBE SOLUTION & CuSO4 solution. FORMED
1. TEST FOR NUCLEOPROTEINS

• Biuret Test
• The formation of purple colored product due
to the reduction of Cu2+ to Cu+ made by the
detected reducing sugar.
• Reagents: NaOH and CuSO4
1. TEST FOR NUCLEOPROTEINS

• Purple or Violet solution

 HYDROLYSIS

• Chemical process involving the addition of

water causing a molecule to cleave into two
parts Phosphate
Nucleic Acids
• Nucleoprotein Nucleotides
Protein

D-ribose
Pyrimidine
Purine
 2. MILD ACID HYDROLYSIS

REMAINING ADD 20 mL PERFORM THE NOTE THE

FILTRATE IN A OF 10% FOLLOWING COLOR
BEAKER H2SO4 TESTS FORMED
 2A. TEST FOR THE PRESENCE OF
PHOSPHATES

1 mL OF THE ADD 2 mL OF
ADD 2 mL OF 5% AMMONIUM HEAT TO
TEST
HNO3 AND MOLYBDATE BOILING
SOLUTION SOLUTION
2A. TEST FOR THE PRESENCE OF
PHOSPHATES

• Ammonium Molybdate Test

• Hydrolysis of pyrophosphate to phosphate
forming yellow precipitate
• Reagent: Ammonium Molybdate, 4 H2O,
HNO3
2A. TEST FOR THE PRESENCE OF
PHOSPHATES

• Light yellow solution with

yellow precipitate
 2B. TEST FOR THE PRESENCE OF
RIBOSE

1 mL SOLUTION FROM
THE ACID HYDROLYSIS

ADD 3 mL OF PLACE TESTUBE IN

BIAL’S ORCINOL BOILING WATER
1 mL OF 0.1% RIBOSE REAGENT TO EACH BATH UNTIL A
SOLUTION TEST TUBE COLOR DEVELOPS

1 mL OF 0.1% GLUCOSE
SOLUTION
2B. TEST FOR THE PRESENCE OF
RIBOSE

• Bial’s test
• Test reagent dehydrates pentose to form
furfural. Furfural further reacts with orcinol
and the iron present in the reagent produce a
bluish product.
• Reagent: concentrted HCL, orcinol, ferric
chloride
2B. TEST FOR THE PRESENCE OF
RIBOSE

• Filtrate: Brown solution with

Black precipitate
• Ribose: Dark Brown solution
with Black precipitate
• Glucose: Dark Brown solution
with Black precipitate
2C. TEST FOR THE PRESENCE OF
PURINES

ADD 3 mL OF 10%
MIX 2-3 DROPS OF
NH4OH TO 2 mL
5% AgNO3
OF THE SOLUTION
SOLUTION TO IT.
IN A TEST TUBE.
2C. TEST FOR THE PRESENCE OF
PURINES

• Principle: Hydrolysis of N- β glycosidic

bonds between purine bases and
ribose/deoxyribose results in a release of
purine bases (A and G) caused by NH4OH.
Ag+ precipitate caused the formation of
cloudy solution.

• Reagents: AgNO3 and NH4OH

2C. TEST FOR THE PRESENCE OF
PURINES

• Cloudy solution with white

precipitate
WHAT IS THE EFFECT OF THE NUMBER
OF A-T OR G-C BASE PAIRS ON THE
MELTING POINT OF DNA?
• A-T Base pairs consist only of Adenine and Thymine
which melts at approximately 69ºC
• Only 2 H+ bonds
• G-C Base pairs consist only of Guanine and Cytosine
which melts at approximately 110ºC
• Only 3 H+ bonds
• The higher the number of G-C base pairs the higher the
temperature needed to melt
• Tm= 4 (G+C) +2 (A+T)
ENUMERATE THE COMPONENTS OF THE
MONONUCLEOTIDE POLYMERS OF DNA
AND RNA

• 5-carbon sugar → deoxyribose and

ribose
• Phosphate group
• Nitrogenous base
GIVE THE METHOD USED FOR THE
QUANTITATIVE ASSAY OF DNA IN A
GIVEN SAMPLE.

• Polymerase Chain Reaction (PCR)