Names: Marielle P.

Balo Date Performed:

Kristine Jade D. Bertulano Date Submitted:

Exercise No. 9
Isolation of Yeast RNA

I. Objectives of the experiment

 To isolate yeast RNA

 To know the purpose of each step and reagent used in RNA isolation
 To explain the purpose of each reagent used in RNA isolation
 To perform recrystallization of RNA

II. Glassware/ Reagents/ Materials Used

 Beaker  Centrifuge
 1% NaOH  Centrifuge tube
 Distilled water  Plastic pipette
 Dry yeast  Litmus paper
 Water bath  95% ethyl alcohol
 Stirring rod  Conc. HCl
 Cheesecloth or wire gauze  Thermometer

III. Schematic Diagram of Procedures

Dilute 2.5mL of 1% NaOH

If turbid, centrifuge
+ 12.5 mL water Evaporate the supernate
or filter and over a water bath to
+ 1.5g dry yeast decant. approximately 5mL.
in a beaker.

Add cooled supernate

Heat on a water bath for
+ glacial acetic acid If necessary, centrifuge
15min with occassional
until faintly acidic to or filter and decant.
stirring.
litmus.

Strain through Allow to cool to 40°C or

Centrifuge
cheesecloth or gauze. lower.
Continuation…

Pour the remaining layer

with vigorous stirring into Air dry.
10mL of 95% ethyl Weigh.
alcohol
+ conc. HCl.

Allow the RNA to settle in Wash the residue twice Calculate the percentage
a covered vessel. with ether. of RNA obtained.

Wash the residue twice

Decant.
with 95% ethanol.

Recrystallization of RNA (optional):

Dissolve RNA in enough Wash the residue twice

1% NaOH. with ether. .
Filter

Faintly acidify with Wash the residue twice

Allow to drain.
glacial acetic acid. with 95% ethyl alcohol.

Pour the remaining layer with

vigorous stirring into Dry at room
0.5mL of 95% ethyl alcohol
Decant.
temperature.
+ 0.005mL conc. HCl.

Transfer into a small

Allow to settle
covered test tube.

IV. Data and Results