Re: [DIYbio] Re: Designing a DIY Gene Electroporator

Maybe we can learn from nature:
Lightning‐triggered electroporation and electrofusion as possible
contributors to natural HGT among prokaryotes
http://arxiv.org/ftp/arxiv/papers/1212/1212.5436.pdf

On Tue, Nov 5, 2013 at 8:36 AM, Cathal Garvey
<cathalgarvey@cathalgarvey.me> wrote:
> In response to "is there a standard wave?", I'll just jump in and say:
> though I've barely ever worked with electroporation, and know not what
> I say, I used to work with those who did. And their impression, from
> the inside, was that no: there is no standard waveform that will give
> useful, if suboptimal, results across the board.
>
> Waveform varied pretty widely even between cell types extracted from
> the same species, and some strains were just too 'ard to EP without
> serious work to optimise, whereas others would accept DNA if you even
> threatened to buzz them a little.
>
> That said, some thoughts:
> A) EP would never have taken off if there weren't a wide margin for
> error, because without knowing in advance the waveforms for this or
> that species, no successes would have been likely. So some lassitude is
> likely among species that are worth turning to EP to transform, to
> begin with, whereas with those that don't respond without careful
> optimisation, perhaps it's better to either await empirical results and
> custom-build an EPorator, or turn to another method of transformation.
>
> B) Piezos and other "dirty" electroporators might suggest that a
> chaotic or noisy source offers the best route to general-purpose
> transformation. That is:
> - Species tend to be picky about response to particular waveforms
> - Too much electricity is the chief killer when EPorating
> - Efficiency at the correct waveform is *probably* scalar with
> duration, implying that the most important bit is getting *any* time
> at the right frequency, with everything else being a bonus.
> So, perhaps rather than seeking a one-true-waveform, it might be worth
> investigating a waveform that scales from one frequency to another,
> covering everything in between, within the timeframe that delivers a
> generally-accepted-as-OK current load on the cell suspension. So if the
> cell responds to any of the intermediate frequencies or waveforms,
> it'll get at least a few instants of it?
>
> Just some thoughts, poorly grounded in experience. :)
>
> Cathal
>
> On Sun, 3 Nov 2013 12:15:27 -0500
> "Meredith L. Patterson" <clonearmy@gmail.com> wrote:
>
>> Adding TQ to this discussion.
>>
>> Cheers,
>> --mlp
>>
>>
>> On Sun, Nov 3, 2013 at 6:43 AM, Nathan McCorkle <nmz787@gmail.com>
>> wrote:
>>
>> > So with a few capacitors like this (microwave capacitor, 2kV 1uF) in
>> > parallel (capacitance adds in parallel):
>> >
>> > http://www.ebay.com/itm/BiCai-1-05uF-HV-Microwave-Capacitor-CH85-21105-2100V-AC-New-/171157909970?pt=LH_DefaultDomain_0&hash=item27d9ce71d2
>> >
>> > or $1.75 each in packs of 20:
>> >
>> > http://www.aliexpress.com/item/Free-Shipping-Imported-microwave-high-voltage-capacitor/1077977860.html
>> >
>> > would this $7 7kV power supply be useful, could you drop the voltage
>> > with a divider to get 1.8kV?
>> >
>> > http://www.aliexpress.com/item/New-DC-3V-to-7KV-7000V-High-voltage-Generator-Boost-Step-up-Power-Module-free-shipping/1326494805.html
>> >
>> > Then seems like all you'd need is an arduino, a gate driver IC, and
>> > a MOSFET or 3.
>> >
>> > Are these big clunky caps really the 2013 best-fit part?
>> >



--
-Nathan

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