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DD-RT-PCR Eluting Bands From Gels

Helge Weissig helgew at ljcrf.edu
Tue Jul 5 21:28:11 EST 1994


In article <2vctr2$q3k at news.iastate.edu>, John Fagan <jfagan at miu.edu> wrote:

> Dear DD-RT-PCR Aficionados,
> We are not having great luck eluting DD products from dried gels.  What
> are other!s experience with this?  If you are successful with this, what
> are the specifics of your procedure?  Is it significantly better to work
> with undried gels?  If so, have you shifted to labeling with 32P, or is
> 35S still adequate?
>         John Fagan

I generally dry my gels under vacuum at 80oC, cut the bands out and elute
by incubation in 100ul TE at 98oC for 10 min.. Never had any problems
(after 50 eluted and reamplified bands!)

good luck,
Helge

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Helge WEISSIG                              e-mail: helgew at ljcrf.edu
La Jolla Cancer Research Foundation        phone: (619) 455 6480 x253
10901 N. Torrey Pines Rd.                  FAX: (619) 455 0181
La Jolla, CA 92037                         
U.S.A.
         
     God invented surfing, so that the really talented people
     would not rule the world. - unknown



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