More Spacing Questions
Tue Apr 22 10:25:15 EST 1997
IHG <Steve Mullen>; mullen at lenti.med.umn.edu wrote in article
<5itkkj$otb at net.bio.net>...
> Lately I have been having trouble with out ABI373 machine being able to
> space properly. Many of the lanes end up with the infamous -12. The
> control has been hit-and-miss, although if I reanalyze I can usually get
> it to space properly. When I look at the gel image it looks pretty good,
> although the bands don't appear to be resolving perfectly--they look
> somewhat smeared. When I look at the analyzed window the bands appear to
> be spaced evenly and the signal strength is fine. This problem seems to
> have come on just in the past couple of weeks. My client reactions appear
> identical to the control, but of course they often can't be coaxed
We use a 377 but I guess this should apply to 373's as well...
The smeary gels you describe may be a result of a build-up of gunk on the
plates. This makes the gel bind less well to the plates and causes a loss
in resolution. One thing that helps is if you wipe the gel side of the
plates with 0.2M NaOH before rinsing of with distilled water. I would also
stear clear of tap water and any cleaning agent such as alconox. We wash
our plates in distilled water, rinse off with milli-q and then leave to
dry. This improved matters for us almost instantly and we now get much
better read lengths.
Hope this helps.
Animal Health Trust
PO Box 5
+44 0638 661111
eryder at hgmp.mrc.ac.uk
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