ARGH! ****GC RICH AREA********

Calvino Ka-wing Cheng ckwcheng at acs4.acs.ucalgary.ca
Mon Jun 10 22:52:39 EST 1996


Hello!

	I was wondering if you could help me.  I'm doing PCR on a
plasmid in the attempt to "site specifically" mutagenise the
resulting enzyme.  The residue which we picked is exactly in the
middle of a GC rich region which encodes for it.

1.  Using PCR, is there a way of unravelling the GC rich primer
(56% GC)?  Can we heat it up?

2.  We are using blunt end ligation.  Is there another way of
creating a primer in a different spot and then using sticky end
ligation?  If so, could you advise us.

Any suggestions?  If you want clarification, e-mail me.  Thanks
for your anticipated help.

Calvino



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