Dominik Haudenschild August 10, 1993 at CBRC

1) Phenol:Chloroform extract

2) Chloroform extract

3) Add: EDTA to 5mM

Tris to 10mM (pH 8.0)

SDS to 0.5%

Proteinase K to 50ug/ml

4) Incubate 30 minutes at 37'C

5) Inactivate Proteinase for 10 minutes at 68'C

6) Extract with Phenol:Chloroform

7) Extract with Chloroform

8) Ethanol precipitate

9) Phosphorylate with PNK if primers are going to be ligated

10) Run on 1.1% low melting Agarose to gel purify

Reference: Nucleic Acids Research, Vol. 19 No. 1, 1991

Reported 20 fold increase of good ligations over non-Proteinase

treated controls.