Any good reviews for gel-shift analyses.

Martin Leach leach at mbcrr.harvard.edu
Mon Sep 27 04:10:03 EST 1993


Hi netters,

looking for some decent literature on performing gel-shift analyses. Have
performed a number using protocols from current protocols, and a butchered
protocol taken from Pharmacia gel-shift kit manual.

Does anyone have any good literature/reviews for trouble shooting
gel-shifts and/or performing them.

My problem seems to be what I think is non-specific DNA binding, even in
presence of 20ug of poly di-dc. This band does not even disappear when I
add as much specific competitor as possiblie (up to 300 ng to 10000cpm
probe).

Tried using a smaller probe (40bp) and now it seems there is less
non-specific interactions but now I have to use 20ug or more of nuclear
extract.

Can all you gel-shift experts out there help a newbie in this field

Many thanx in advance

Martin Leach.

Please feel free to e-mail me at leach at mbcrr.harvard.edu or just follow-up
this article.

A summary of comments will be posted.



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