HYDROLYSIS OF

NUCLEIC ACIDS
Group 8

Objectives
To
To

be able to prepare a hydrolyzed and unhydrolyzed DNA and RNA

be able to perform qualitative tests to determine the

characteristics of nucleic acid present

THEORIES

Hydrolysis
Breaking
Site

of a bond in a molecule using water.

of cleavage: sugar, phosphate backbone, or in the base

Result:

purine and pyrimidine bases, oligonucleotide, nucleosides,

ribose or deoxyribose, and phosphate.

Base-catalyzed hydrolysis
Occurs in RNA
Hydroxyl ion assist a
nucleophilic attack on
the 2 hydroxyl group
on the phosphorus
yields intermediate
cyclic-2,3phosphonucleosides
that hydrolyze to
nucleoside-2phosphates and
nucleoside-3phosphates
UNSTABLE
Use of water to stabilize
forming 2 or 3
nucleoside

Acid-catalysed Hydrolysis
Causes depurination
Cleaves the purine Nglycosyl bonds
Adenine and guanine
are liberated and
apurinic site remains.
Some phosphodiester
bond are cleaved
Much harsher and
acidic is required to
remove pyrimidine Nglycosides

QUALITATIVE TEST
DATA

DNA
Test

Control

DNA

Acid
Hydrolysate

Base
Hydrolysate

Benedicts Test

Sky blue

Sky blue

Sky blue

Sky blue

Orcinol Test

Rusty brown

Light brown

Yellow brown

Deoxyribose

White soln
with brown ppt

Red soln with

brown ppt

White soln
with brown ppt.

Purine bases

Cloudy
lavender to
clear solution
with white ppt

Clear solution
with White
precipitate

Clear soln with

brown ppt

Uracil and
Cytosine

clear solution

More cloudy

Bright yellow
to clear
solution(Ba(OH
)2

Biuret Test

Light blue

Light blue

Light blue

Xanthoproteic

Light yellow

Clear solution

Light yellow

RNA
Test

RNA

Acid Hydrolysate

Base Hydrolysate

Benedicts Test

Sky blue

Apple green

Sky blue

Orcinol Test

Dark bue-green

Dark green with

black particles
solution

Dark blue-green

Deoxyribose

White cloudy with

brown ppt

Red solution with

brown ppt

Layer of orange,
white and brown
solutionwith brown
ppt.

Purine bases

Cloudy dirty white

soln

Light pink soln with

brown ppt

Clear soln with black

ppt

Uracil and Cytosine

More cloudy

Cloudy white

cloudy white

Biuret Test

Light purple

Light purple

Light blue

Xanthoproteic Test

Bright yellow

Light yellow

Bright yellow

Inorganic Phosphate

sample

BenedictsTest

Orcinol test

Test for deoxyribose

Test for Purine bases

Test for uracil and cytosine

Biuret test

Xanthoproteic test

ANSWERS TO
QUESTIONS

How can intact DNA be obtained

from a solution of DNA and RNA
Dna

could be obtained from a solution of DNA and

RNA by using a base-catalyzed hydrolysis, since in
alkaline solution RNA is destabilized forming
nucleosides while the DNA remains stable and
intact.

Based on the qualitative test, How can you

differentiate hydrolyzed from unhydrolyzed RNA
Hydrolized

Rna is positive for orcinol test, test for purine

bases, biuret test, xanthoproteic test.

-acid-catalalyzed hydrolysis: presence of pentoses,

pyrimidines, and
phosphoric acid
- base-catalysed hydrolysis: only for pentoses and
pyrimidines.
Unhydrolized

Rna is only positive for phosphate test.

Based on this experimental result, do you think

you obtained pure RNA from yeast? Why or Why
not?
We

were not able to obtain pure Rna from yeast because of the
positive result from our xanthoproteic and biuret tests

Based on these experimental result, do you think

you obtained pure DNA from onion? Why or why
not?
We

were not able to obtain pure DNA because of the positive test
of xanthoproteic on our DNA samples.

CONCLUSION

 Base-catalysed

hydrolysis occurs only in RNA

Acid

hydrolysis of nucleic acids yeilds purines, pyrimidine bases

and phosphoric acid

Despite

the errors in the hydrolysis and qualitative test experiment

we have understood the nucleic acid hydrolysis

Reference:
http://

www.scribd.com/doc/38672208/Hydrolysis-of-Nucleic-Acids#scribd

http://

www.chem.uky.edu/courses/che554/5_MolBio/NucleicAcidTechnology
IntroChpt.pdf