to assess the purity of nucleic acids.

For pure phosphate or a crust of phospo-ammonium

RNA A260/280 is ~2. molybdate.

Alkaline Hydrolysis s a chemical process in The test for Purines or Murexide test gives the
which a molecule is cleaved into two parts by the reaction of RNA with nitric acid given that purines
addition of a molecule of water in a solution with are known to be readily soluble in dilute acids.
a pH greater than 7. In cellular systems, Yellow precipitate upon evaporation resulted from
the phosphodiester bond between the oxidation process by Concentrated HNO3.
two ribonucleotides can be broken by Nevertheless, it turned red when moistened with
alkaline hydrolysis because of the free 2' a base, which is a positive result for presence of
hydroxyl group. The stability and lifetime of purine bases (guanine or adenine).
RNA is affected by alkaline hydrolysis. Base
hydrolysis of RNA extracts pentoses, purine In the test for pyrimidines, which is a
bases, proteins, and phosphoric acid. qualitative test for the pyrimidine bases cytosine
and uracil, which produces a green coloration
Table 2. Chemical Characterization for the when the sample is treated with bromine water.
Hydrolyzed RNA and Standard Solutions The addition of barium hydroxide will turn the
liquid purple.
RNA from RNA from
Chemical Yeast Yeast Standard
Test (theoretic (experimen solutions
al results) tal results) REFERENCE
colorless blue green [1]The Department of Biochemistry, Faculty of
Test for dark green Pharmacy, University of Santo Tomas, Laboratory
solution with sol’n
Ribose sol'n
white ppt. Manual in General Biochemistry (2009).
yellow yellow layer yellow [2]http://www.scribd.com/doc/25162486/RNA-
Test for
crystalline layer Purification-and-Analysis
Phosphate
ppt. [3]http://www.mercksource.com/pp/us/cns/cns_
reddish reddish hl_dorlands_split.jsp?
Test for
brown brown yellow soln pg=/ppdocs/us/common/dorlands/dorland/eight/
Purines
residue residue 000107483.htm
clear soln [4]http://www.rowan.edu/biology/Labs/biomole.
Test for
violet ppt. with white violet ppt. pdf
Pyrimidines
ppt.
[5]http://www.saddleback.edu/faculty/steh/bio3a
folder/bio3alabmolecules.pdf
During the experimental period, some of the [6]http://library.med.utah.edu/NetBiochem/pupy
results that were obtained did not match with the r/pp.htm
theoretical results. Some factors were taken into [7]http://www.mun.ca/biochem/courses/3107/Le
consideration that maybe the sample was ctures/Topics/bases_and_chains.html
contaminated, or there was lacking of the heating [8]http://www.odec.ca/projects/2004/mcgo4s0/p
during the process. ublic_html/t3/RNA.html
[9]http://answers.yahoo.com/question/index?
Characterization tests are used to describe the qid=20070822220829AAHcPsd
reactions of RNA to different reagents in order to
represent its structural features.
From the book:
For the Ribose Test, the orcin reaction is a test [1]The Department of Biochemistry, Faculty of
for the presence of ribose in RNA. The reaction Pharmacy, University of Santo Tomas, Laboratory
depends on the conversion of ribose to an Manual in General Biochemistry (2009).
aromatic aldehyde (furfural) which then reacts
with Orcin (3,5-dihydroxy toluene) to give a blue- From the internet:
green color.
[2]http://www.scribd.com/doc/25162486/RNA-
For the test for the presence of phosphate in the Purification-and-Analysis
hydrolyzed RNA, the positive result is a yellow [3]http://www.mercksource.com/pp/us/cns/cns_
precipitate. The positive result is because of the hl_dorlands_split.jsp?
reaction of ammonium molybdate solution, which pg=/ppdocs/us/common/dorlands/dorland/eight/
when dropped onto the sample will result to a 000107483.htm
yellow stain which indicates the presence of
[4]http://www.rowan.edu/biology/Labs/biomole.
pdf
[5]http://www.saddleback.edu/faculty/steh/bio3a
folder/bio3alabmolecules.pdf
[6]http://library.med.utah.edu/NetBiochem/pupy
r/pp.htm
[7]http://www.mun.ca/biochem/courses/3107/Le
ctures/Topics/bases_and_chains.html
[8]http://www.odec.ca/projects/2004/mcgo4s0/p
ublic_html/t3/RNA.html
[9]http://answers.yahoo.com/question/index?
qid=20070822220829AAHcPsd