ability to pcr introns from cDNA

Bernhard Mayr ndxdbmay at rrzn-user.uni-hannover.de
Sat Apr 27 07:24:55 EST 1996


Nancy Duncan <duncanna at ohsu.edu> wrote:
>I was wondering if anyone has an opinion on the ability to pcr introns 
>from a cDNA prep...I understand that cDNA by definition is exon only, but 
>is it possible to isolate some pre-mRNA, and then reverse transcribe that 
>to cDNA, which could lead to pcr amplification of introns? We hope the 
>answer is no, but are unsure....thanks....
>

Igues there must be some un- or patially spliced RNA in a good prep,
but I'd worry most about DNA contamination when doing PCR on cDNA.
If there is un- or patially spliced RNA I'd expect it to be at very low 
copy numbers and even DNAse treated RNA might contain low numbers of DNA.

So what would be really diagnostic, is a PCR-product of partially spliced 
RNA species containing 2 or more intron/exon boundaries with some of the 
introns already spliced out.

Bernhard Mayr
Dept Clin Endocrinology
Medizinische Hochschule Hannover, Germany




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