EXPERIMENT 8.

1
AIM: To isolate DNA from available plant material such as
spinach leaves, green
papaya etc. pea seeds,

REQUIREMENTS
Plant material (such as spinach leaves, green pea seeds
green papaya), mortar and pestle,
or
beakers, test tubes, liquid detergent, non-iodised sodium
chloride, distilled water, meat tenderizer or
papain solution/juice of papaya/pine apple juice, 95%
ethanol, spool etc.
PREPARATION OF SOLUTIONS
Detergent salt solution is prepared by adding 10 mL
sodium chloride to 90 mL of distilled water. liquid detergent and 10 g of non-iodised
Meat tenderizer solution is
water (Juice of
prepared by adding 5 gof tenderizer (enzyme) to 95 mL of distilled
papaya/pine apple, filtered through muslin cloth can be used as substitute for
meat tenderizer).
5% NaCl solution is prepared by dissolving 5 g of non-iodised sodium chloride in
distilled water. 100 mL of
Chilling of ethanol must be done by keeping 95% ethanol
in plastic bottle in the freezer over
night.
Core Experiments 55

PROCEDURE
Take 5 g of the plant tissue (spinach leaf/green pea seed/green papaya) and grind it in the
mortar by adding 10 mlL detergent, salt solution and filter it through muslin cloth.
Take 10 mL of the filtrate, add 3-4 mL tenderizer/papaya juice and swirl the test tube by holding
the tube between the two handsthe contents.
to mix

Pour 10 mL chilled ethanol carefully down the side of test tube to form a layer on the top of
the content; let it stand undisturbed for about 3 minutes.
Dsing the glass rod stir gently through interface of the two layers to collect the precipitate of
DNA and place it in a test tube with 5% NaCl or distilled water.
The quantity of DNA present in the given plant material can be estimated through

spectrophotometer.

Fig. 8.1. DNA that separates out can be removed by spooling (spool = reel for winding yarn).

OBSERVATION
The addition of ethanol to the solution causes DNA to precipitation. The DNA fibres appears as
white precipitate of very fine threads on the glass spool.

PRECAUTIONS
1. The plant material should be washed throughly with distilled water to remove any dust and
dried by blotting beforeweighing
All the glasswares used must be thoroughly cleaned and dried.

3. The chemicals and enzymes used for the experiment must be of standard quality which should
be manufactured by standard pharmaceuticals.