Wizard PCR purification from agarose gel

Markku.M Jeltsch jeltsch at csc.fi
Sat Mar 9 03:01:24 EST 1996


On 8 Mar 1996, ruth e.w. walker wrote:

> I am unable to purify my PCR product from the gel using the Wizard kit. 
> It works well when I purify directly from the PCR rxn. I use 2% agarose, 
> which melts within 4 min at 65C in presence of the resin. Is that too 
> long? Too short? What else could be wrong, given that it does work with 
> the PCR rxn, done concurrently?
> 
I stopped using the Wizard kit for PCR purification, because of this and
other problems. Instead I use Qiaex beads (with a slightly modified 
protocol, that includes a 70% EtOH washing step after the two washings with 
the buffer that is provided with the kit). If you still want to use 
Wizard I suggest to reduce the %age of your gel if possible to 1.5. That 
should be enough even for 150 bp fragments. Or try it with low melting 
point agarose.
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Markku Michael Jeltsch           MOLECULAR/CANCER BIOLOGY LABORATORY
Apollonkatu 19A14                THE HAARTMAN INSTITUTE
FIN-00100 Helsinki               UNIVERSITY OF HELSINKI
e-mail: jeltsch at cypress.csc.fi   P.O.B. 21 (HAARTMANINKATU 3)
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       -358-0-454 3418 (home)
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