help lambda-gt11 PCR

Shiao Y. Wang sywang at whale.st.usm.edu
Tue Jun 21 23:04:14 EST 1994


Lois C. Johnson (ljohnson at udcemail.udc.upenn.edu) 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




More information about the Methods mailing list