Hello!!
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
