Taq inactivation

John Lum lumdicks at netvigator.com
Tue Jul 18 02:50:01 EST 2000


Martin Chan wrote:

>     I am going to directly use the PCR mixture for restriction digestion,
> but I fear the remaining Taq in the PCR mixture will fill in the 5' overhang
> generated by restriction endonucleases. Is my worry necessary? Can Taq be
> inactivated by heating at 94-96C for 30 min prior to restriction digestion?
> I am frustrated by the low recovery of my lab's PCR purification kit, so I
> plan to skip the purification step.

        Besides the fill-in problem of Taq, how about the reaction buffer? Is it
compatible with your restriction digestion? I remember Boehringer Mannheim has
compiled a list for the compatible RE with the PCR buffer. Hope this can help.

Regards,
John Lum






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