Phone: 403-782-8100 ext. 268
Fax: 403-782-6120
johnsm@em.agr.ca
>>> Katheryn Resing <Katheryn.Resing@Colorado.EDU> 05/17 3:08 AM >>>
We couldn't get good results with a constant current focusing. We use the
programable power supply from pharmacia, at constant voltage, but varying
during the focusing in steps. I have included below the program we use,
with the instrument settings (amps and watts are limits, not constant
settings).
As long as this is up for discussion, I have a query for the other
users. We have had problems getting reproducible focusing. Strips treated
identically and focused at the same time will show nice, tight spots on
most of the strips, but about 1 out of 4 will have streaking on the higher
mass spots that looks like they haven't focused well.
Any one found a solution to that one, yet?
I've just started making my own IPG strips to solve the problem of inconsistent strips and focusing. I have run about a dozen 2D gels so far and all gels show consistent spot quality and pattern. The spot quality and pattern of gels using labmade strips are the same (maybe even a little better) as the commercial strips. It only takes about 2 hours to set up and pour the gel for IPG strips. One IPG slab can make about 30 strips (at about 50 cents per strip). Cutting the strips precisely is still a challenge.
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