Sequencing Gel Fixation

John Cooper Picard at nih.gov
Sun Nov 20 12:59:33 EST 1994


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



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