Monday, June 07, 2004

Description
This protocol describes a method to precipitate DNA in order to concentrate the DNA fragment or plasmid fo further downstream processes.

Procedure
1 Extract DNA fragment following PCR reaction using a commercial gel purification kit.

2 Precipitate by adding 0.1 volume of 3M NaOAc (pH 5.2) followed by 2 volumes of 100% ethanol. Put at -20 degrees for 30 mins.

3 Centrifuge sample at 12,000 g for 10 mins at 4 degrees. 30 mins if lower DNA concentration expected (e.g. <100 ng/ml).

4 Remove supernatant. add 200 ul of 70 % ehtanol. Centrifuge at 12,000 g for 5 mins at 4 degrees.

5 Remove supernatant. Air dry pellet for approximately 10 mins.

6 Resuspend in TE (pH 8).

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