RT-PCR

Jonathan Hecht jhecht at ucsd.edu
Thu Apr 28 10:49:24 EST 1994


In article <leach-260494212126 at ppp-8-28.bu.edu> Martin Leach,
leach at mbcrr.harvard.edu writes:
>BTW if doing RT-PCR - DNAse treat your RNA before RTing it. Otherwise
>unless u r pcring over an intron (in the DNA) u won't know whether the
>product is real or due to DNA contam.

As well DNAse treating the RNA, its important to do a control PCR
reaction using mock-reverse transcribed RNA as a template (do this by
adding everything to the RNA except reverse transcriptase).  That way you
will be absolutely sure you have no DNA contamination.  For a good
reference about this, see the SRY1 cloning paper (Goodfellow, I think).

Jonathan



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