purify PCR products using Wizard Prep (Promega)
Robert Solomon Bioc
rgs at mole.bio.cam.ac.uk
Wed Jun 15 03:45:51 EST 1994
In article <CrE9Gu.Cx7 at acsu.buffalo.edu> vp23qw83 at ubvms.cc.buffalo.edu (Xin Guo) writes:
>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.
>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).
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