help lambda-gt11 PCR

Shiao Y. Wang sywang at
Tue Jun 21 23:04:14 EST 1994

Lois C. Johnson (ljohnson at wrote:
: I was wondering if anyone knows whether or not PCR can be done with lambda
: gt11 or gt10 DNA or plate lysate.  I have tried this procedure twice with

I amplify DNA inserts about 1.5 kb in lambda gt22A routinely. I use the
normal protocol of the AmpliTaq pol. I remove a plug from the plate
contain the plaque of interest and soak it in 50 ul TE for several hours. I
use 1 ul of the liquid in a 50 ul PCR reaction. It's very straight forward
thus I'm surprised it's not working for you. I would check that the
primers you are using are suitable (sounds too simple to be the problem, I
know). I use the standard gt11 forward and reverse primers. The sequences are
in catalogs from companies like Promega.

Good luck.

Shiao Wang
University of Southern Mississippi

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