ammonium persulfate in sequencing gels

Bill Melchior wmelchior at ntet.nctr.fda.gov
Wed Mar 17 16:09:53 EST 1993


All instructions I have seen for sequencing gels call for adding ammonium
persulfate as a 10% solution.  For several years, I have merely added an
equivalent amount of dry powder to the acrylamide/bis/urea solution just
before degassing, and it seems to work fine (for manual 35S sequencing). (I  
use 0.5 mg ammonium persulfate per ml gel solution and then, after
degassing, add 0.5 microliter TEMED per ml.)  This saves the extra step of
dissolving the persulfate. 

My question is, why do the procedures call for adding a persulfate 
SOLUTION?  And in particular, is it necessary when preparing gels for 
fluorescence sequencing machines, which in general seem to be more 
demanding regarding gel quality?
________________________________________________________________________________
Views expressed are not necessarily those of NCTR, its sponsoring agencies,
or the United States government.

Bill Melchior                                ||       MISSING LINK
National Center for Toxicological Research   ||    Man's a kind
Jefferson, AR  72079                         ||    of Missing Link,
(501) 543-7206                               ||    fondly thinking
                                             ||    he can think.
WMELCHIOR at NTET.NCTR.FDA.GOV                  ||       from Grooks, by Piet Hein



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