I have been using the same settings which are preset in the instrument. The
arcing stopped after limiting the volume of ligated mix to 1/20th the volume
of Electrocompetent cells. Initially I was adding 2microlit ligated mix for
20microlit comp cells, now I'm adding only 1microlit.
Thanks for all the suggestions!
--Yg
> Message: 1
> Date: Mon, 02 Feb 2009 11:27:21 GMT
> From: Aawara Chowdhury <aawara from pontiff-playground.org>
> Subject: Re: Electroporation
> To: methods from net.bio.net> Message-ID: <tMAhl.12549$%j5.8911 from newsfe10.iad>
> Content-Type: text/plain; charset=us-ascii
>> In <6unrddFg2336U1 from mid.individual.net>,
> Christian Praetorius <prae from gmx.net> wrote:
>> > Aawara Chowdhury <aawara from pontiff-playground.org> wrote:
> >
> >>Oddly enough this has failed for me - many, many, times. We use 1 mm gap
> >>cuvettes, and 1800 volts (25 uF capacitance, and I think 200 ohms
> >>resistance). What are your settings?
> >
> > I used the same settings, when we did electroporation. As long as the
> > cells were washed good enough, we never had problems with short
> > circuits. Since we have a quite good protocol for making
> > supercompetent cells, we moved away from it.
>> Can you share this protocol with us?
>> Thanks!
> AC
>> --
> Email: echo 36434455860060025978157675027927670979097959886449930P | dc
------------------------------
>> Message: 2
> Date: Mon, 02 Feb 2009 09:06:53 +0000
> From: Christian Praetorius <prae from gmx.net>
> Subject: Re: Electroporation
> To: methods from net.bio.net> Message-ID: <6unrddFg2336U1 from mid.individual.net>
> Content-Type: text/plain; charset=us-ascii
>> Aawara Chowdhury <aawara from pontiff-playground.org> wrote:
>> >Oddly enough this has failed for me - many, many, times. We use 1 mm gap
> >cuvettes, and 1800 volts (25 uF capacitance, and I think 200 ohms
> >resistance). What are your settings?
>> I used the same settings, when we did electroporation. As long as the
> cells were washed good enough, we never had problems with short
> circuits. Since we have a quite good protocol for making
> supercompetent cells, we moved away from it.
>> Christian
>> --
> X-no-Sig: yes
>>> ------------------------------
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