[Open_electroporator] Pragmatic electroporation protocols

[Keoni Gandall]

Hey

I was rushing around over the weekend trying to keep electrocompetent cells
cold when prepping them, and ended up just getting a whole lot of arcing
(which is pretty scary, there is a large flash and pop, with the
electroporation cassette is for the most part destroyed).

Room temperature prep would be nice, so I was going to test that this week,
but I decided to do some investigating. Apparently, electrocompetent cells
can be prepared at room temperature-
http://www.nature.com/articles/srep24648

The reason why (hypothetically) is explained pretty well here-
https://www.reddit.com/r/askscience/comments/33n3w9/why_do_we_need_to_keep_competent_cells_on_ice/

Essentially, the hypothesis is that the cold makes lipids in the membrane
seal slower. When doing heat shock, which is pretty slow, this is likely
helpful.

They propose in the nature paper that ice-cold temperatures change the
membrane topology and survivability. Since the mechanism of competence is
the electric field, (while with heat-shock it appears to be ions and
temperature) which is applied quickly and sometimes lethally, the slow-down
that cold provides likely doesn't outweigh the survivability costs, as more
healthy cells will survive electroporation.



Overall, if you are making your own electrocompetent cells, perhaps try to
do it at room temperature instead of rushing around with ice. I know that's
wayyyyy easier for me in a DIY setting.

-Keoni
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