pcr overlap extension technique

Magdalena Pleschberger mpleschb at edv1.boku.ac.at
Fri Feb 23 04:50:30 EST 2001


Fellow bio.netters
I am trying to do an gene fusion based on the overlap extension pcr
technique.

I'm able to get the  two separate pcr fragments (with the overlapping
region ) but the
pcr with the flanking primer for the complete product (+4200 bp) doesn't
work.


maybe an unfortunate  circumstance is also, that the two
fragments do have a different GC concentration (the longer
fragment:38% the shorter one 58%).
it would be great to hear from you and the way you solved that
problem,
best wishes and thank you very much for your time,
sincerely
Magdalena




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