Help with RT-PCR!!! What's wrong with my primers?

Duncan Clark blackhole at abuse.plus.com
Fri Jul 23 07:50:01 EST 2004


Historians believe that in newspost 
<356e5246.0407230439.5bfa6405 at posting.google.com> on Fri, 23 Jul 2004, 
Miao <meng.meng at plantphys.umu.se> penned the following literary 
masterpiece:
>I even tried those primers to amplity from plasmid (containing my
>gene's cDNA), but none of them worked!


If they won't amplify from the plasmid then either the cDNA you think 
you have isn't, the primers are wrong (or wrongly labelled) or you have 
a something else wrong with the PCR.

Have you tested the PCR reagents with a known working set of primers and 
shown that those reagents work OK.

If the cDNA is correct then there is little point running a more 
expensive RT-PCR until you can get primers that PCR on that cDNA plasmid 
template.

Duncan
-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.



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