deaerate or not deaerate

Richard P. Grant see_sig at cmtech.co.delete.uk
Thu Aug 20 10:38:27 EST 1998


In article <6rhb7a$7ep$1 at info.cyf-kr.edu.pl>, "Zbigniew Rudzki"
<mprudzki at cyf-kr.edu.pl> wrote:

: Does degasing of polyacrylamide belong to the group of totally magic and
: irrational procedures contaminating molecular biology ?
: Today, forced by the technical reasons, I tried for the first time to skip
: it and I added twice as much TEMED as usually (20 uL per 70 mL).  It
: resulted in an almost instant catastrophic polymerization :)
: 
: So what is your opinion on degasing, after all rather troublesome a
: procedure ?

I understood that O2 inhibits polymerization, therefore after you've had
the acrylamide a while it makes sense to degas.  However, I hardly ever
de-gassed, maybe I just used it too quickly, but the one or two times I
found that polymerization times were increasing (and the AMPS was < 1
month old) de-gassing did help, a little, for large sequencing gels.

When I was trying to get some really nice mini-protein gels, and wanted
wells formed all the way to the top of the gel, I allowed polymerization
to occur under N2.  Seemed to help, too.

R

-- 
Richard P. Grant MA DPhil     |             rgrant at cmtech.co.uk
Senior R&D Scientist          |             work: www.cmtech.co.uk
Cambridge Molecular           |      home: www.avnet.co.uk/adastra           
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