DNA sequencing gels

KARN at Butler.EDU KARN at Butler.EDU
Mon Jul 19 15:54:50 EST 1993

Dear Sequencing Netters:
	I have relatively little experience running DNA sequencing gels and
am having problems with samples spreading across other lanes when I am loading
them.  I use a BRL S2 gel of the standard 0.4 mm thickness and the usual
concentrations of gel components and urea.  It seems that I have this problem
more often (but by no means soley) when I pour the gel the night before and
run it the next day (I have been led to believe that this practice is accep-
table if one takes measures to keep the gel from drying out while sitting
overnight).  The spreading problem does not seem to be due to the shark's
tooth comb penetrating the gel to any particular degree.  Sometimes I get
very good loading (i.e. no spreading across lanes) and sometimes gross cross-
lane contamination - and I can see no pattern to why it works well sometimes
and not others.  I would be most grateful for tips from you oldtimers in the
sequencing game.
Bob Karn

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