jxie at magnus.acs.ohio-state.edu
Thu Apr 28 23:19:44 EST 1994
In article <2polu4$ajr at network.ucsd.edu>,
Jonathan Hecht <jhecht at ucsd.edu> wrote:
>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).
I generally include a negative control in my RT rxn in which I add
RNAase A treated RNA instead. You should not get anything from this
control when pcring. If you do :-( ===> DNA contamination!!!!!
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