random priming for RACE, not polyT

Peter Ellis pjie2 at cam.ac.uk
Wed Mar 24 03:20:32 EST 2004


domo11711 at yahoo.de wrote:
>The problem for RACE and all the kits is:
>The only way to reach 5'-end, is to go from 3'-end, from polyA. And
>that is the problem, because some of mRNA (or RNA) are very long. To
>use a specific primer on mRNA makes no sense, it will prime
>everywhere. 

Glad you told me that, you've just chucked years of science out the 
window.  All those people doing RT-PCR with gene-specific primers must 
just be wrong, I guess.

>
>Because RNA has higher affinity, and even 6 bp is enough
>for priming.
>To get first strand ssDNA, random priming could solve this problem.

So... you think gene-specific primers aren't specific enough, so you 
want to use random primers.  Care to run that one by me again?

Peter



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