cDNA synthesis and linear amplification!
csakis at ki.se
Mon Feb 7 05:04:19 EST 2000
I am wondering if anybody tried to amplify the amount of mRNA during the
first strand cDNA synthesis. The idea is very simple. You use oligo(dT) for
priming, do the synthesis, then heat the sample to 80-85 degree, (maybe 94)
and let the sample cool down and add reverse transcriptase again. This way
you double the amount of mRNA you had in the beginning. If you do it again,
you triple your mRNA. In some application you need very high amount of
polyA+ RNA, then this would solve the problem, and you do not introduce PCR
generated bias or artefacts.
Did anybody try this method?
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