RT-PCR

Theo Thijs theo.thijs at med.kuleuven.ac.be
Mon Jun 22 07:53:46 EST 1998


It depends on the reverse transcriptase you used to synthesis your ss-cDNA.
For instance if you use SuperScripte II RTase the reaction temperature
should be around 42 degree C. If you use AMV, it can stand for 55 degree C
which favors for the overcoming second structure in your mRNA template. Hope
this helps.

Elena Ayaso wrote in message <358E5879.2F0D at ucg.ie>...
>I am using oligo dT to do reverse transcription on RNA from embryos.
>Then I use specific primers on the 1st strand cDNA I get no PCR product.
>When I use specific primers to do the reverse trnscription it works.
>I think the oligo dT might not be working. I would like to know what
>anneling tempeture I should use if I want to use 18mer oligo dT as a
>primer to check that it's O.K.
>Elena Ayaso
>Thanks





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