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)
phone: -358-0-434 6436 (work) FIN-00014 HELSINKI
-358-0-454 3418 (home)
fax: -358-0-434 6448 (work)
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