Inverse PCR

Viraj Master vam2 at midway.uchicago.edu
Wed Oct 12 10:51:10 EST 1994


In article <37gioc$pak at mserv1.dl.ac.uk>, chealy at hgmp.mrc.ac.uk (Dr. C.P.
Healy) wrote:

> I want to design an inverse PCR experiment to isolate the 5' regions of a number
> of transcripts which are differently spliced. I have a number of references to
> start from but would like to hear the experiences in others. The main things th   
> I am worried about are the efficiency of circularisation of the cDNA (how dueo you
> ensure blunt endedness and what are the optimum conditions for intramolecular
> ligation) and how do I ensure efficient amplification of  long cDNA fragments
> (e.g. 2-3kb - do I need to use one of the new Taq preparations?). Please reply
> by e-mail - I'll post if there's enough interest.
              ************************************

Chris, please do post a summary.

Thanks,

Viraj

-- 
Viraj Master
Dept. of Organismal Biology and Anatomy
University of Chicago



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