Sequencing Gel Fixation

Wiggy A.C.Hilton at bham.ac.uk
Mon Nov 21 13:07:38 EST 1994


In article <Picard.57.785354373 at nih.gov>, Picard at nih.gov (John Cooper) wrote:

> In article <CzF6MI.58r at demon.co.uk> Anthony Tomlinson
<tomlinson at pplros.demon.co.uk> writes:
> >From: Anthony Tomlinson <tomlinson at pplros.demon.co.uk>
> >Subject: Re: Sequencing Gel Fixation
> >Date: Thu, 17 Nov 1994 16:18:18 GMT
> >In article <tsute.267.000B56F6 at microbio.umass.edu> Tsute Chen,
> >tsute at microbio.umass.edu writes:
> >>I've heard that the fixation step (with 10% acetic acid & 10% methanol)
> >can be 
> >>skipped without much difference in the result during the normal Sanger's
> >DNA 
> >>sequencing procedures. Does anyone have any experience or opinion about
> >this?
> >
> >I remember that some people said they had problems in
> >humid environments with unfixed gels, but most seemed
> >to feel that fixing sould be omitted if you're in a hurry.
> > I've not had the nerve yet!
>  Not fixing works best with modified acrylamides like Long Ranger...
> Here at NIH they are trying to decrease the amount of mixed radioactive 
> waste so we use Long Ranger and don't fix. As long as the temp of the gell 
> stays under 50c, the gels look great.
> 
> John
> 
> John Cooper, MD
> National Inst. of Health
> Picard at helix.nih.gov

I always used to fix my sequencing gels but recently I have omitted this
step without any problems.
Wiggy.



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