[BiO BB] Re: max mA in 2D-DIGE

[Jean-Etienne Poirrier]

Bart,

Are you sure of your solutions? You didn't say
anything about your gel concentration. Once, we made a
mistake in the gels concentrations and it went (a lot)
faster.

We are not considering the current when running our
2nd dimension gels but we are considering power ...
Here is our protocol:
Step 1. 2.5W per gel during 30 minutes (maximum of
600V or 400mA)
Step 2. 100W for all gels for a maximum of 5 hours
(maximum of 600V or 400mA ; I think it's 5W for all
gels for an overnight run but I don't have the
protocol here).
What are your other parameters?

Bart, I'm also a university student in Belgium but at
the ULg and working with 2D-DIGE. If you have any
other questions, feel free to ask.

Yours,
Jean-Etienne

--- Bart.Boerjan at student.kuleuven.be wrote:
> Date: Thu, 12 Oct 2006 21:00:15 +0200
> From: "Bart Boerjan"
> <Bart.Boerjan at student.kuleuven.be>
> Subject: [BiO BB] max mA in 2D-DIGE
> To: <bio_bulletin_board at bioinformatics.org>
> 
> dear colleague scientists,
> 
> I'm a university student in Belgium at the KUL. I'm
> working on proteomics 
> now and one of the techniques I'm using is 2D-DIGE.
> 
> I would like to run a strip in second dimension
> loaded (via IEF) with 
> bacterial cellular peptides. normally we adjust the
> mA so that the gel runs 
> for about 13 hours. this afternoon we tried for the
> first time 72mA (instead 
> of 24) for a tank with two gels (so 36mA per gel)
> and after two hours the 
> tracking dye was already half of the gel, so i
> wondered could a seperation 
> that goes this fast cauze complications?
> 
> I found his mailing list via google search, and I
> hope maybe one of you can 
> answer me, or share an experience
> 
> sincerely
> 
> Bart Boerjan


Jean-Etienne Poirrier, je64p at yahoo.com
http://www.poirrier.be/~jean-etienne/
http://www.epot.org/blog/

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