Polyacrylamide gel electrophoresis query
Timothy R Jackson
tjackson at uoguelph.ca
Tue Apr 8 14:34:26 EST 1997
we have recently noticed in our lab that a significant amount of, what
appears to be, precipitation of small fragments (<300bp) out of our gels
onto the glass plate that they're fixing on. our fragments are 33p
labelled and we've noticed that the longer you fix a gel the worse the
problem (hotter plate than gel!) a 0.4mm gel only needs to fix for a
minimum of 20min which does not cause much of a problem but fixing
overnight, for instance, seems to result in massive precipitation. we
are getting away from fixing all together - the drying time is slightly
longer and no difference in the results (and less waste too!!)
Tim Jackson
Dept. of Zoology
University of Guelph
Guelph, Ontario
CANADA
: I was wondering if anyone could give me a concise answer to a
: query I have regarding polyacrylamide gel electrophoresis
: methodology.
: The question is this ; when fixing gels with acetic acid &
: methanol, what is happening ? (How does this fix
: oligonucleotide bands ? By precipitation ? & are small oligos
: likely to be leached out during this process ?).
: Many thanks in advance for your help.
: Mark.
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