Poor capillary transfer?
Lab_Mac_Hanson at QMrelay.mail.cornell.edu
Fri Mar 18 12:53:24 EST 1994
In article <2mcfbeINNib7 at newsstand.cit.cornell.edu>, douglas l feinstein
<dlfeins at cumc.cornell.edu> wrote:
> Until we get our vacuum blotter, we are transfering our northerns
> using 10xSSC capillary transfer overnite. But what we are finding is that
> even after 16 hr transfer, only the 18S band is transfering, while the
> higher stuff is staying very nicely in the gel. I have made sure that
> there are enough paper towels and enough buffer in the reservoir. I was
> thinking that maybe the weight I put on top of the paper towels is too
> heavy, thus crushing the gel and preventing transfer of HMW bands. Any
> other ideas?
I have recently had good luck with *downward* transfer of RNA, much better
than upward. S&S (no endorsement) sells a kit that might make it less
clumsy to set up As far as your question about mashing your gel, yes,
rumor has it that as the gel collapses (with weight) nucleic acids are more
likely to get trapped, and of course large RNAs are transferring out more
slowly than small RNAs.
Hello from "the other Cornell"
Claudia Sutton, cas9 at cornell.edu
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