GAPDH PCR: DNA vs. RNA...
rhubner at gins.uia.ac.be
Thu Jul 9 08:56:32 EST 1998
I checked a DNA sample for amplification potential by PCR with GAPDH
primer pair... This primer pair spans
intron "H" [for 4205-4225; rev 4742-4761] and is frequently used to
control for DNA contamination when setting
up RT-PCRs [RNA gives 450bp whereas DNA gives 550bp]...
I am puzzled because I observe _ONLY_ _450bp_ in this extract... The
DNA was extracted from cell line SVEC
(SV40 transformed endothelial cells)... Extract on gel shows neat HMW band
and no obvious RNA contamination; this was PCR with Taq polymerase and
manual hot start...
Can an intron get "lost" from entire genome?
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