[Open_electroporator] Pragmatic electroporation protocols

[William Beeson]

What type of water were you using.  I would make sure to use distilled
water or better and not tap water.

On Mon, Jun 27, 2016 at 5:21 PM, Keoni Gandall <koeng101 at gmail.com> wrote:

> 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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