Hello, netters. I have a high-secondary structure (high gc content,
anyway) mRNA that I am trying to do RT-PCR on, and I haven't been
successful in reading through the region with oligodT or gene-specific
primers and SuperscriptII/ RNaseH kit, following the kit protocol, using
ice-quench and no ice-quench after RNA melting, using a variety of first
strand synthesis temps. (42,50,55). I am currently trying random
hexamers, and the next step if that doesn't work is the more thermostable
So, has anyone tried the new Thermoscript RT (BRL)? anyone have
preferences re. Tth vs. Thermoscript? Tips for "difficult" transcripts?
phillinj at slu.edu
St. Louis University Hospital
3635 Vista Ave.
St. Louis MO 63110 USA