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14. dissolve 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.