1.

2.
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4.
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6.
7. .
8. .
9.
10.
11. .
12.
13.
14. the pellet in distilled water or TE.
15. Measure the concentration by OD at 260nm and 280 nm or by diphenylamine reaction.
16. Check the DNA by running on 0.8% agarose gel electrophoresis.
Requirements
1. Lysis buffer
Components: NaCl 150mM
TrisHCl 20mM (pH 8)
EDTA - 50mM
SDS - 0.5%
Preparation: mix 15 ml of 1 M Nacl, 2ml of 1M tris (pH- 8), 10 ml of disodium EDTA
(0.5M) and 2.5 ml of 20% SDS and makeup the volume 100ml.
To be noted:
a) Disodium EDTA does not dissolve until the pH is 8. So dissolve the EDTA by
adding NaOH pellet. At point of pH 8 it will dissolve
b) Do not autoclave SDS. SDS will dissolve automatically on standing. Store at
room temperature.
2. TE buffer : TrisHCl - 10mM (pH 8)
(Tris buffer)
EDTA - 1mM

Preparation: Mix together 1ml tris buffer (pH 8) and 200 l of EDTA 0.5 M and make
up the volume to 100ml.
3. Buffered phenol l(water saturated phenol is sufficient)
4. Chloroform : isoamyl alcohol (24:1)
5. 3M sodium acetate (pH 5.2, 3M pH adjusted with glacial acetic acid)
6. Absolute alcohol
7. 70% ethanol
Result
Genomic DNA is isolated and run on 0.8% agarose gel electrophoresis, visualized and
photographed.