PCR product purifn. for sequencing

Wayne Woods wgwoods at ariel.ucs.unimelb.EDU.AU
Mon Jul 4 19:09:52 EST 1994


In article <9407042125.4e17d561.LN1 at lincoln.cri.nz>,
Scrimshaw, Brian J <SCRIMSHAWB%GRIEHFS at LINCOLN.CRI.NZ> wrote:
>Purification of PCR products for sequencing.
>
>I have sequenced lots of PCR products manually in the past, and
>have tried lots of different ways of purifying PCR products prior
>to sequencing.
>My method of choice has been to use promega magic (now wizard)
>preps. They are fast real easy and give excellent results for PCR
>products in the 300-600bp range.
>I am currently trying to purify a 118bp band for sequencing. At
>this size the wizard prep has a very poor yield after gel
>purification (just done it, zip on the gel). Can someone give me
>advice of a good way to purify a band this size? I'm most

Brian,

Are you using the normal Wizard minipreps, or the 'Wizard PCR Preps'
kit? I've had nothing but consistently *good* results using the latter. 
I isolate 1 kb - 1.5 kb PCR bands from normal (high mp) 1 % agarose,
and get a satisfactory % recovery (never actually worked out the % though).

Wayne

________________________________
Wayne Woods
Department of Microbiology
University of Melbourne
wgwoods at ariel.ucs.unimelb.edu.au
________________________________



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