longer primer?

Elliot Goldstein e.goldstein at asu.edu
Thu Jun 29 16:35:16 EST 2000


we are having the same problem with the 5' end of our gene. if it doesn't fall
out soon with standard PCR and nuclease protection we will try one of the
advanced RACE kits. i have seen 2 others besides clontech, invitrogen and
ambion.
if anyone has any other ideas or insights into which race kit is best, i would
like to know.
thanks,
       elliott goldstein

Gregor Bucher wrote:

> HI Jamie,
>
> seems that you are trying a kind of RACE. In the (quiete expensive) kits
> you ligate an adaptor to the mRNA. Than you have a good primer upstream
> and downstream. Unfortunately it es not that straightforward as you read
> it here - I have tried various mRNAs and some do work, some don´t. The
> newer (and probably better) kits do not ligate blunt but take advantage of
> a special RT-Polymerase that adds polyG (orC?) at the end. One address is
> clontech (there are cheaper ones but I don´t remember the names)
>
> Hope this helps!
>
> Gregor
> --
> Gregor Bucher, Zoologisches Institut der LMU München
> 0049/89/5902/444
> st2153 at zi.biologie.uni-muenchen.de
> ----------------------------------

--
Elliott S. Goldstein
Biology Department
Molecular and Cellular Biology Program
Arizona State University
Tempe, AZ 85287-1501
480-965-7176
e.goldstein at asu.edu








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