??IBI-UEA electroelutor
Michael SZardenings
msz at chemie.fu-berlin.de
Fri Apr 21 09:55:35 EST 1995
These electroeluters work fine only in theory, but one can do a few
things to improve yields. Most people will loose the DNA on removal from
the apparatus, so I usually did the following:
-use 3M NaAcetate in the slot, that contains some stain (!) like
bromophenol or xylenecyanol (just as agarose loading buffer). Best to use
both, as they move differently. After the run try to remove the layer on
top of the stains + some stain, that's were the DNA is.
-perhaps use a tip that fits into the hole, but does not enter too much.
This allows you to empty the slot completely.
-try this first without DNA to get some practice. The stain allows you to
run the elution to a maximum time, as you can follow the stain moving
through the high salt buffer cushion, with the DNA probably on top of it.
-spot <1myl loading buffer or stain solution in front of the gel slice, this
gives extra help when estimating completeness of the elution.
Still, I never got really great yields either, but 40-50% should be
possible without problems.
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