Can I increase RT stringency?
phelan at jsei.ucla.edu
Thu Sep 28 14:17:35 EST 1995
I have been using RT-PCR to amplify a member of a group of closely
related sequences. The system is such that the forward primer
probably exactly matches more than one of these sequences, and
so I need to rely on the reverse primer to generate the specificity.
Unfortunately, I end up with bands from more than one sequence.
I think this is primarily due to mispriming at the RT step.
For my use, I really want only one band. Therefore,
my question is this:
Is there a way to increase the stringency during RT with MuLV to
prevent mispriming? I would prefer to make adjustments without
switching enzyme systems.
Please reply by e-mail, as I can't always reach the news group.
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