PCR from phage?

Markus Piotrowski Markus.Piotrowski at rz.ruhr-uni-bochum.de
Mon Nov 20 18:06:25 EST 1995


bonhamp at duke.usask.ca (Peta C. Bonham-Smith) wrote:
>Hi:
>Has anyone been able to PCR amplify an insert directly from phage
>particles as oppposed to isolating phage DNA first?  Do we need to treat
>the phage prior to PCR?
>
>Peta Bonham-Smith
>Assistant Professor
>University of Saskatchewan
>
>bonhamp at duke.usask.ca

Dear Peta,

I did PCR with degenerate primers and a lambdaZAP-cDNA-bank. The only
thing to do is to 'denaturate' the phages for 10 min at 95°C and maybe to
rise the amount of the primers. I usually took 2 ul of the cDNA-bank and
400 pmol of each (degenerate) primer in a final volume of 50 µl. Hope
this will help you, good luck

Markus




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