Electrophorating with used cuvettes

ww40 William_D_WARREN at UMAIL.UMD.EDU
Fri Nov 15 15:43:00 EST 1991


G'day to JJW, John Nash, Paul Fisher and everyone else out there
who has been on about reusing electrophorator cuvettes (and why is
it that so many ozzies have views on this...well here's another's).

We have been reusing electrophorator cuvettes for some time now
and havn't had any problems. Since >90% of the time we use
electrophoration to rescue plasmids passaged through
insect embryos, we have had to develop a rather thorough procedure
to guard against cross contamination. Immediately after use, cuvettes
are thoroughly rinsed in H20 and left to soak. At the end of the day
we then incubate them overnight at 37oC in a solution of 50mM Tris,
10mM MnCl2 and 2ug/ml of DNase. Cuvettes are then washed extensively in
H20 and then left in 100% EtOH. Just before use we air dry them to remove
the EtOH, but I guess that an 80oC dry sterilization would probably
be better.

Since starting to reuse cuvettes we always include a no DNA control,
and on only a few occasions have we seen a colony, but none
looked like coli.

For general transformation purposes though I'd recommend one of the
procedures already posted (using DNAse is a double edged sword).

***************************************************************************
 BILL WARREN             "Due to the current recession the light at the
 Ctr. Ag. Biotech             end of the tunnel will be turned off....
 Univ of Maryland                       until further notice"
***************************************************************************




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