Activity No.

10
ISOLATION OF SOLUBLE RIBONUCLEIC ACID

Soluble ribonucleic acid is a mixture of species ranging in molecular weight from 25,000 to
35,000 (75 to 100) nucleotides). RNA can be isolated from baker’s yeast by heating with alkali
which extracts nucleic acids and water soluble proteins and inactivates nucleases which can
degrade RNA. The nucleic acid is then separated from associated proteins and other
interfering substances by acid extraction at pH 4-5. The final step is treatment with alcohol
acidified with concentrated HCl to precipitate the RNA followed by repeated washing with
alcohol and ether or other organic solvents to remove lipids.

MATERIALS

balance thermometer plastic spoon

beaker, 250 mL, 500 mL (2) graduated cylinder, 10 mL
Bunsen burner litmus paper
centrifuge machine stirring rod clay
triangle test tubes, 10-mL (6)
evaporating dish test tube rack
filter paper test tube brush
water bath wire gauze

CHEMICALS

acetic acid, glacial alcohol (95%) ether hydrochloric acid, conc.

sodium hydroxide (1%) yeast

PROCEDURE

1. In a beaker dilute 5.0 mL of 1% NaOH solution with 25 mL water.

2. Add 10 grams of dry yeast. Heat on a water bath for 15 minutes with occasional
stirring.
3. Centrifuge at high speed for 10 minutes. Decant and to the supernatant add glacial
acetic acid one drop at a time until faintly acidic to litmus. Pour the acidified
supernatant with vigorous stirring into a 250-mL beaker containing 20 mL of 95% ethyl
alcohol with 0.2 mL of conc. HCl. This treatment will precipitate the RNA.
4. Allow the RNA to settle. (See Appendix: Common Laboratory Techniques, Fig. C.1a
and C.1b).
5. Decant and wash the RNA twice with 1mL of 95% ethanol and twice with ether.
6. Allow to air dry until the next lab period.
7. After drying, show the RNA to your instructor.
8. Reserve for Activity No. 17 “Hydrolysis of Nucleic Acids”.
 Activity No. 10
ISOLATION OF SOLUBLE RIBONUCLEIC ACID
OBJECTIVE

DATA/OBSERVATIONS

1. Show the RNA to your instructor: (10 pts)

Source :

Youtube Video
provided .

2. Describe the RNA obtained: (4 pts)

a. color:

b. texture:

QUESTIONS

1. Give the purpose of the following reagents in each step involved in the isolation
procedure performed. (8 pts)

a. alkali

b. acid
 c. acidified alcohol

d. ether

2. The following is an incomplete schematic diagram for the isolation procedure used.
Label each step with the reagents used and products obtained. (16 pts)