purify PCR products using Wizard Prep (Promega)
dbradley at vax1.tcd.ie
Wed Jun 15 06:53:40 EST 1994
In article <2tmf3v$4ma at lyra.csx.cam.ac.uk>, rgs at mole.bio.cam.ac.uk (Robert
Solomon (Bioc)) wrote:
> In article <CrE9Gu.Cx7 at acsu.buffalo.edu> vp23qw83 at ubvms.cc.buffalo.edu (Xin Guo) writes:
> >Hi, Bionetters:
> >I have a trouble to get a high yield of PCR products by using Promega's
> >Wizard Prep method. Instead of using lower melting agrose gel, we use
> >normal agarose to seperate the PCR products, cut the band, and boil
> >it to melt the agarose. Then I follow the Promega's steps to mix the
> >DNA products with the resin, add it into the column, and elute it.
> >It is said the recovery can be >90%, but we only get <10%. Are there
> >any of you has ever utilized this method to purify DNA from agarose gel?
> >Any suggestion will be great appreciated.
> >Xin Guo
> >SUNY at Buffalo
> >Department of Pharmacology and Toxicology
> >vp23qw83 at ubvms.cc.buffalo.edu
> I think this is another example of what, in years to come, will be
> known as ... The Promega Effect ... <cue sinister music>
> There seems to be batch-dependent (in)efficiency in Wizard PCR Preps,
> or some variable not taken into account by the protocol - use one
> of the glass milk methods (no, no vested interest here, just bitter
> experience - standard disclaimer applies) or freeze-squeeze (yes, I'd
> rather use phenol and purify some DNA than use a kit and lose it).
As a fellow victim of the Promega Effect - I am interested to find out what
exactly "freeze squeeze" is and how good a purification technique it is.
Phenol would definitely be the lesser of two evils.
EMAIL dbradley at vax1.tcd.ie
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