[DIYbio] Re: Designing a DIY Gene Electroporator

Nathan McCorkle nmz787 at gmail.com
Sun Nov 3 23:58:23 UTC 2013


On Sun, Nov 3, 2013 at 9:14 AM, Meredith L. Patterson
<clonearmy at gmail.com> wrote:
> Is there a standard reference, or much work in the literature, on pulse
> shape/duration/&c? I've seen several papers that essentially boiled down to
> "we empirically determined the most effective settings on our Bio-Rad
> electroporator for transforming $ORGANISM_WE'RE_INTERESTED_IN", but what is
> it that makes square waves so great?
>
> (My boyfriend the electrical engineer is the one asking this -- it'd be a
> lot easier for him to build a capacitor bank where the tail of each pulse
> trails off, and take fewer parts, but will that reduce the number of
> transformants?)
>
> Cheers,
> --mlp

This isn't specific for DNA transformation, it's talking about
chemotherapy molecules too, but it looks pretty similar to the other
stuff I've seen (and it does mention DNA quite a bit):
http://lbk.fe.uni-lj.si/pdfs/beme2004.pdf

otherwise there's this book (I've got it in paperback, I'll leaf
through it later or tomorrow):
http://www.sciencedirect.com/science/book/9780080917276

this looks OK, not much on actual devices:
http://www.physics.uoguelph.ca/~dutcher/download/handbook%20of%20biological%20physics/18.pdf

Looks like bipolar pulses can give higher transformation at lower
amplitude (voltage). One thing mentioned Electrofusion can be started
by a long-ish DC pulse, then a few AC pulses... or a DC pulse followed
by AC dielectrophoresis (the electrodes in dielectrophoresis are
covered in a dielectric insulator). Not sure how much lower AC pulses
can be than DC though...


this is quick and cheap, but some key points to refresh:
http://www.harvardapparatus.com/hapdfs/HAI_DOCCAT_3/W3_9_356.pdf


I'm sure there are more, I don't have better articles handy but there
should be some more in the DIYbio archive. See the attached paper as
well.

I can't work on this anymore today, but I'll search my papers this
week for sure.

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