RT-PCR in GC-rich Chlamy

Julie Knott jknott at biosci.cbs.umn.edu
Wed Dec 18 07:55:09 EST 1996


I am looking for advice from anyone doing RT reactions on our GC-rich
Chlamy!! I have performed RT reactions with both random hexamers and gene
specific primers using Superscript II RT (GibcoBRL) with successful
results in all but one region.  I have complete genomic sequence in
this region and can somewhat predict the cDNA sequence, (the RT-PCR will
confirm it).  The problem region appears to have at least one 40bp region
of 82% GC!  I assume that the RT reaches that region and cannot proceed.

Has anyone tried performing RT reactions for the purpose of RT-PCR
or RACE on very high %GC templates?  What methods did you use to keep the
RNA denatured?

Thanks in advance,

Julie Knott (Porter Lab)
jknott at biosci.cbs.umn.edu

More information about the Chlamy mailing list

Send comments to us at biosci-help [At] net.bio.net