EXPERIMENT 9

I S O L AT I O N O F Y E A S T R N A
Name: Irene B. Velado
Group number: 1

Date: October 26, 2015

Section: 1 TF 1-4 PM

ISOLATION OF YEAST RNA

Table 9.1. Observations for the isolation of RNA from yeast.
Mass of yeast (g): 1.5039
Appearance of yeast

golden yellowish and small

Addition of 1% NaOH

Milky-brown solution with a soya milk like smell

Heating

Lighter milky-brown solution but more viscous

Appearance of the supernatant

liquid

Clear yellowish solution

Addition of glacial acetic acid

Still clear yellowish solution (faintly acidic)

Appearance upon cooling and

addition of 95% ethanol

Turbid solution. Formation of precipitation

Appearance of the residue

Powdery white precipitate

1. Why is the yeast used as the source of RNA?

Yeast is used as the source of RNA because the ratio RNA-DNA is the highest in these
microorganisms. Moreover, the fermentation and recovery techniques for yeasts are well
established. Also, it is inexpensive and readily available.
2. What is the purpose of the following in RNA extraction?
a. NaOH
NaOH increases the pH level of the solution resulting in the denaturation of contaminant
proteins, inactivate nucleases which can degrade RNA.
b. Heating
Heating helps loosen the cell membrane by increasing the kinetic energy of the lipid
molecules, making it release more RNA.

isolation of yeast rna

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c.

Centrifuging

Centrifugation, which uses the idea of gravity, breaks up the cells and leads to the
sedimentation of the largest particles present in the nuclei. Increasing the centrifugal speed
brings about the sedimentation of first the cytoplasmic large and small granules. RNA was
obtained from the cytoplasmic fractions. After centrifugation the supernatant was collected.
d. Glacial acetic acid
The purpose of adding glacial acetic acid is to lower the pH level to help denature more
proteins, prevent alkali RNA hydrolysis and to ensure the desired RNA will not degrade.
e. Cooling to -10oC or lower
The purpose of cooling to -10oC or lower is to prevent RNA degradation. It limits RNases
activity since this proteins are very stable and can withstand high temperature and present
everywhere.

References
Elson, D. 1965. Metabolism of nucleic acids (macromolecular DNA and RNA. Annu. Rev. Biochem
34:449-86
Stryer, L.; Berg, J.; Tymoczko, J. 2007. Biochemistry. San Francisco: W.H. Freeman
Biochemistry Laboratory Manual, Institute of Chemistry, UP Diliman, 2007 Edition. pp. 10-15.

isolation of yeast rna

| Chem 145.1 Department. of Chemistry UP Visayas \ jbrojo