Long RT-PCR

hetherington at mbcf.stjude.org hetherington at mbcf.stjude.org
Thu Jun 2 12:56:18 EST 1994


We have routinely done RT-PCR of 2,000 bp.  ANother 600 bp shouldn't be a
problem.  If you are amplifying eukaryotic products, try starting with an
oligo-dT primer, and MMLV reverse transcriptase.  Extension times should be
about 1 min per 1,000 bp.  No experience with bacterial genes, but you would
need to use a specific primer.  In either case, use the whole RT reaction
(10ul) for the amplification step.



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