Difficult RT-PCR

Dr. D. Zha zha at mpi-muelheim.mpg.de
Wed Oct 27 10:41:53 EST 1999


How did you do your RT, I mean what kind of primer in your RT, if you
used oligu(T), you may use random primer just for trying. Good!



Andrea Gilpin wrote:
> 
> Dear Everyone:
> 
> We are currently facing difficulty getting RT-PCR products.  Our gene
> has a genomic length of 9 kb, in which there are 8 exons making the
> total coding sequence of 3.6 kb.  We can easily get a 1.2 kb cDNA
> spanning exons 1 and 2, and recently, we have also obtained a 2.9 kb
> fragment of cDNA from exon 1 to exon 6.  But we have had tremendous
> difficulty getting the full-length cDNA with the RT-PCR method.  Does
> anyone have any suggestions, recommendations and/or tips.
> 
> Thanks very much,
> 
> Zhou
> zhou at botany.utoronto.ca



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