Electroporation

yoginee budhkar via methods%40net.bio.net (by eenigoy from gmail.com)
Wed Feb 4 05:01:09 EST 2009


 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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