Re; plate electroporation device
coady at ere.umontreal.ca
Sat Nov 9 00:29:17 EST 1991
In response to a couple of requests for the reference regarding
the plate electroporation device that I wrote in about, check
Biotechniques Vol. *, No. 1 (1990) p. 28 (or thereabouts),
"A Simple and Effective Electroporation Apparatus", by
J.F. Speyer. He later published an addendum in the same journal,
but I don't have a copy of that. I believe that it dealt with
electroporating multiple samples by using a larger capacitor. The
only drawback that I had with this system was getting the distance
between the two steel plates set up properly. I found that a layer
of Scotch tape worked, provided that it was three pieces thick. Any
more and the efficiency plummeted.
For those who are afraid of electrical devices or who need
calcium shock to introduce "salty" DNA into their bacteria, Inoue et
al published a nice article in Gene (vol. 96, pp. 23-28) where they
describe a few changes to the Hanahan procedure for competent cells,
principally a low temperature for growing the cells. They got
transformation efficiencies equivalent to what they could get from
electroporation and (this is the big payoff here) they got 1-3 x
10 to the 9th transformants per ug using DNA up to 200 pg. And that
is a ton, compared to what most published efficiencies are tested at.
May not be important to some, but for making a cDNA library you've
gotta play these angles :-). Mike Coady (COADY at ERE.UMONTREAL.CA).
P.S. I just noticed that I screwed up the volume number up above,
and I'm not used to our new Unix system (i.e. can't correct text).
It should read Vol. 8.
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