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Biology Practical - Experiment No. 11

1. The document describes an experiment to isolate DNA from plant materials like spinach leaves, green pea seeds, and papaya. 2. The procedure involves grinding the plant tissue, filtering it with a detergent-salt solution, adding a tenderizing enzyme or fruit juice, and then precipitating the DNA by adding chilled ethanol. 3. When ethanol is added, DNA precipitates out of the solution and appears as a white precipitate of fine threads that can be collected on a glass rod or spool.

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Siddhant Singh
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3K views2 pages

Biology Practical - Experiment No. 11

1. The document describes an experiment to isolate DNA from plant materials like spinach leaves, green pea seeds, and papaya. 2. The procedure involves grinding the plant tissue, filtering it with a detergent-salt solution, adding a tenderizing enzyme or fruit juice, and then precipitating the DNA by adding chilled ethanol. 3. When ethanol is added, DNA precipitates out of the solution and appears as a white precipitate of fine threads that can be collected on a glass rod or spool.

Uploaded by

Siddhant Singh
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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EXPERIMENT 8.

Objective. To isolate DNA from available plant material such a s spinach leaves,
green pea seeds, papaya etc.

REQUIREMENTS
Plant material (such as spinach leaves, green pea seeds or green papaya), mortar and
pestle, 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 liquid detergent and 10 g of non
iodised sodium chloride to 90 mL of distilled water.
is by of
Meat tenderizer solution prepared adding 5g of
of
tenderizer (enzyme) to 95 ml
apple, filtered through muslin cloth can be used as
distilled water Juice papaya/pine
substitute for meat tenderizer).

5% NaCl solution is prepared by dissohving 5 g of non-iodised sodium chloride in 100


mL of distilled water
ethanol must be done by keeping 95% ethanol in plastic bottle in the freerer
Chilling of
over night.

54
cORE EXPERIMENTS

5
PROCEDURE

.Take 5 g of the plant tissue (spinach leaf green pea


the mortar by adding 10 mL detergent, salt solution seed/green papaya) and grind it in
and filter it through muslin cloth.
Take 10 ml of the filtrate, add 3-4 ml
.

by holding the tube between


tenderizer/papaya juice and swirl the test tube
the two hands to mix the
contents.
Pour 10 mL
.
chilled ethanol carefully down the side of test tube to forma layer on the
top of the content; let it stand undisturbed for about 3 minutes.
Using the glass rod stir gently through interface
of the two layers to collect the precipi-
tate 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 fibres
The addition of ethanol to the solution causes DNA to precipitation. The DNA
appears as white precipitate of very fine threads on the glass spoo.

PRECAUTIONS
, The plant material should be washed throughly with distilled water to remove any dust

and dried by blotting before weighing.


All the glasswares used must be thoroughly cleaned and dried.

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

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