pcr overlap extension

Magdalena Pleschberger mpleschb at edv1.boku.ac.at
Fri Feb 23 06:22:22 EST 2001


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 if you have assuggestion how to solve that
problem,
best wishes and thank you very much for your time,
sincerely
Magdalena





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