specific 'nested' priming for reverse transcription in RT-PCR

Soenke Behrends soenke at zedat.fu-berlin.de
Wed Nov 2 14:53:25 EST 1994


 I want to do reverse transcription with a specific
additional downstream oligo. From what I have heard
thermostable enzymes like Tth or Retrotherm don't
work as well as for example Stratascript that I use
routinely. So I habe decided on a 17mer oligo with
an annealing temperature of 40C.
What about RNA concentrations (polyA or total)?
Primer concentrations?
thermal profile?
Does anyone have any experience?
Is it worth the effort?
Can you go further up with the RNA amount when you have
transcripts of low abundance?

Thanks in advance




More information about the Methods mailing list