V The breaking of the DNA double-helix

bonds by the addition of water

V Complete hydrolysis of chromosomal

nucleic acids gives the inorganic
phosphate, 2-deoxyribose and four
different heterocyclic bases: Adenine,
Guanine, Thymine, Cytosine
V !ses strong acid at high temperature to
break down the stable structure of DNA
into its components

V Conditions:
À Acid: 6M HCl
À Temperature: 100° C for 1 hour
V These conditions break both of the
phosphate ester bonds and also the N-
glycosidic bond between the
deoxyribose and the purines and
pyrimidine bases.

V The products of this mixture are the 4

bases, phosphoric acid, and deoxyribose
which then polymerises to produce a
brown sticky tar.
V Monosaccharide
V Present in the DNA structure
V Discovered in 1929 by Phoebus Levine
V ¦t can identify DNA chemically.
V Positive result: blue solution
V Reagents used: diphenylamine reagent
V Principle: conversion of pentose to a
molecule that reacts with the
diphenylamine reagent
V „

 
  


 
      
V Nucleotides are building blocks of
Nucleic acids.
V Three compounds are needed in the
formation of a Nucleotide: deoxyribose;
a base; and a phosphate group.
V They combine through 2 condensation
reactions.
V ëasic structure of a nucleotide:
V Nucleic Acids are formed by the linking
of 2 or more nucleotides by the
formation of Phosphodiester bonds.
V The Phosphodiester bond is formed by
the condensation reaction between the
²OH group of the phosphate group in a
nucleotide and a ²OH group on the
carbohydrate ring of another
nucleotide.
V DNA is formed from 2 polynucleotides.
V ¦t has a double helical structure with
bases on the inside and phosphate
outside of the helix.
V They are held together by the pairing of
nucleotides bases through H-bonding.
V Test for a phosphodiester bond linked at
C3 or C5 in the ribose.
V Positive result: Yellow precipitate
(Phospho-ammonium molybdate)
u
Ñ

V Principle: The Phosphate ions react with
the ammonium molybdate to form a
phosphomolybdate complex.
¦n the test for presence of purines, DNA is
reacted with Nitric acid since Purines are
known to be readily soluble in dilute acid.
Nitric acid oxidized it leaving a yellow
precipitate upon evaporation; however it
turned red when moistened with a base, a
positive result for presence of purine bases
V a

  

 

 
 




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O ë

O ë NH2
HN CH3
HN
HN
O N O N
H H N
O H
!racil Thymine Cytosine
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V Dische Test
À Dehydration of deoxyribose forming hydroxyl
levulinaldehyde
À Complexation reaction with diphenylamine
V Murexide Test
À Oxidation of purine by concentrated HNO3
forming dialuric acid and alloxan
À Condensation reaction leading to formation
of alloxanthan
À Neutralization leading to formation of
murexide or ammonium purpurate
V -heeler Johnson Test
À [ormation of dialuric acid by neutralization

V Phosphate Test
À Addition, Precipitation and Complexation