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INTRODUCTION
Ribonucleic acid (RNA) molecules are singlestranded polymer nucleic acid
made up of
ribonucleotides. Each ribonucleotide consists of a
ribose sugar, a phosphate, and a nitrogenous
base derived from purines which are adenine (A)
and guanine (G) and from pyrimidines which are
cytosine (C) and uracil (U). In comparison with
DNA, RNA is relatively shorter in molecules thus
it is not easily damaged by shearing but it is
more unstable and more prone to degradation
due to having a ribose sugar.
Ribonucleotide Components
Figure 1.
A 260/ A 280
(absorbance) at the
A 260/ A 230
(absorbance)
EXPERIMENTAL
A. Materials
The materials used for isolation of yeast were
active dry yeast (Saccharomyces cerevisiae),
concentrated HCl, glacial acetic acid, 1% NaOH,
95% ethanol and ether. The reagents used for
alkaline hydrolysis and characterization of RNA
were 0.3N NaOH, 1N KOH, concentrated H2SO4,
concentrated HNO3, ammonium molybdate,
bromine water, 10% KOH, Ba(OH)2,orcinol
reagent, and standard solutions : pentose sugar
of RNA (ribose) and nitrogenous bases of RNA
(adenine, guanine, cytosine and uracil).
B. Procedure
1. RNA Isolation from Yeast
The diluted mixture of 5mL of 1% NaOH
solution, 25mL of water and 3.0g of dry yeast
was water bath for 15 min. at 60 degrees Celsius
with occasional stirring. After heating, the
mixture
was
strained
with
cheesecloth,
centrifuged and acidified with glacial acetic acid.
The mixture solution of 20 mL of 95% ethanol
and 0.2 mL of conc. HCl was poured to the
supernatant and stirred vigorously. All residues
were decanted, centrifuged, and washed twice
with 2mL of 95% ethanol and with ether.
2. Alkaline Hydrolysis
The mixture solution of 2 ml of 0.3N NaOH
isolated RNA was water bath for an hour.
hydrolyzate was cooled and adjusted to pH
by using glacial acetic acid using pH paper.
hydrolysed sample RNA was used for testing.
and
The
4-6
The
1.
was
Chemical
Test
Std.
solution
RNA
from
yeast
(+)
result
Test for
Ribose
Dark
Green
solution
Green
solution
Green
solution
Test for
Purines
Yellow ppt
Yellowish
brown
ppt
Reddish
brown
ppt
Test for
Pyrimidine
s
White ppt
Litmus:
Red to
Blue
Brown
ppt
Litmus:
Red to
Blue
Purple
ppt
Figure 2.
reagent .
Reaction
of
ribose
with
orcinol
REFERENCES
[1]
Crisostomo, A., et. al. (2010). Labaratory
Manaul in Genearal Biochemistry. Quezon City:
C&E Publishing Inc., pp. 73-77.
[2] Balda, K. , et. al. (2011) Nucleic Acid RNA
[PowerPoint
slides].
Retrieved
from
http://www.slideshare.net/kevbalda/report-exp6-and-7-dna-and-rna
[3] Ribonucleic Acid. (n.d.). In Scitable online.
Retrieved
from
http://www.nature.com/scitable/definition/ribonu
cleic-acid-rna-45