PCR problem with EGFP

Björn Fritz bfritz at ix.urz.uni-heidelberg.de
Mon Sep 20 09:10:08 EST 1999

Hi all,

I have severe problems in doing a PCR with a plasmid, that contains
GFP.  The primers I designed prime in the enhanced GFP gene. I used 4
different primers, checked them with several primer design programs,
used different temperatures, polymerases and buffers, checked the
quality of the template. I am quite sure that the template is okay
because I used as well the original plasmid I got from Clontech. But
all primers don´t work even on the plasmid. Another group working for
years with the same plasmid states that it has the same problems.
Clontech has no idea.
Does anyone of you has  primersequences in the EGFP that work well? I
would be very glad for any suggestions

Thank you

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