Thermoscript v. Tth for RT-PCR

Nancy Phillips, M.D. phillinj at SLU.EDU
Thu Mar 4 10:35:52 EST 1999


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 
RTs. 
	So, has anyone tried the new Thermoscript RT (BRL)? anyone have 
preferences re. Tth vs. Thermoscript? Tips for "difficult" transcripts?
Many thanks! 

Nancy Phillips
phillinj at slu.edu
Pathology
St. Louis University Hospital
3635 Vista Ave.
St. Louis MO 63110 USA
ph:(314)577-8782
fax:(314)268-5120



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