Inverse PCR

Shehrebanoo Malik banoo
Tue Nov 26 22:09:17 EST 1996


hi!  I have done inverse PCR successfully on a genomic DNA template,
but it would be a bit more complicated to get it to work on mRNA.
Actually, since IPCR relies on having a known restriction site between
the flanking (primer) regions of known sequence, you would have to
use cDNA as your template.  

Is your problem that you are not able to amplify an entire transcript
by RT-PCR, so you want to try IPCR?  You might want to try degenerate
gene-specific primers for your first-strand cDNA synthesis, or try 
screeening a cDNA library with your RT-PCR fragment of cDNA as a probe.


Good luck!


Banoo Malik, 
c/o Dr. G. Drouin
Dept of Biology
University of Ottawa
banoo at bio01.bio.uottawa.ca


Andrew Doherty <A.Doherty at Bris.ac.uk> wrote:
>Cristina Juarez wrote:
>> 
>> Dear netters:
>>                 I'm trying to do Inverse PCR from mRNA. Does anybody have
>> a working protocol for it?  Please reply to: cjuarez at omni.cc.purdue.edu.
>> Many thanks!!
>> Cristina
>WHat's Inverse PCR? Do you mean RT-PCR?
>-- 
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>Dept. Anatomy			Tel (0117)9287421
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