Method for Anchored PCR/RACE
Martin Kennedy
cytogen at chmeds.ac.nz
Sun Oct 18 17:14:27 EST 1992
In article <1992Oct15.143546.10299 at magnus.acs.ohio-state.edu>, gchacko at magnus.acs.ohio-state.edu (George W Chacko) writes:
> In article <1992Oct15.162328.123 at chmeds.ac.nz> cytogen at chmeds.ac.nz (Martin Kennedy) writes:
>
>> Finally, anchored PCR doesn't seem to work over regions much longer
>> than about 500bp, at least in my experience. Some PCR reactions show
>> a nice tight band, others a smear -
>
> I've gotten tight bands upto 1 Kb using a 3' RACE system. I use 2 ug
> of total RNA (the quality of RNA appears to be the most critical
> factor IMO).
Sorry - I meant 5' RACE doesn't seem to work over much more than 500bp, but
maybe someone has some comments about that. I guess it is the efficiency of
the RT step that is critical here (and of course the RNA quality)
Cheers
Martin
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