Differential Display PCRs - help needed!
Miss J.M. Cameron
jcameron at hgmp.mrc.ac.uk
Wed Mar 27 13:24:20 EST 1996
I am currently having terrible trouble remamplifying cDNAs that I have cut out from
differential display gels.
My method is to cut out the band from the gel, elute it for an hour in TE and then
boil it for 5 minutes.
I usually try PCRing 1, 2 and 3 microlitres of the eluted DNA in a 20 microlitre
PCR reaction (2.5uM anchor primer, 0.5uM 10mer primer, 20uM dNTPs, Promega Taq and
Promega buffer with Mg2+).
Sometimes these PCRs work, and sometimes they don't. At the moment I can't get
them to work at all!!!
Has anyone had success with this stage of the DDRT process??
Thanks for your help!
Univesity College London, UK
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