Purifying PCR product from 2% agarose gel

[Ednot]

I´ve had the same problem on a Min Elute GEl extraction Kit, this kit
is based on cartridges membrane, if you apply too much
sample or wrong pH, the sample will elute on the wash.

The QiaexII is based on charged silica, this item of the kit is very
labile
for my opinion, once i tried a new silica (newer lot) and it worked
just fine,
Once again check the pH of your sample.

A less partic method is to put your slice of gel with your band in a
eppendorff tube
and some TE buffer and let it a Max RPM on your centrifuge for 4
hours, remove the remaining agarose.
And precipitate the DNA with Ammonium Acetate 7M, pH 8.5 and then
Ethanol 100%.

Good Luck


On Feb 18, 10:46 am, "hilary cassidy" <cass... from gmail.com> wrote:
> Hi
>
> I've been trying to purify my PCR product from a 2% agarose gel all
> week now as it needs to be purified prior to Topo cloning. I've used
> both the QIAEX II Gel Purification kit by Qiagen and the Illustra DNA
> and Gel Band Purification Kit by GE Healthcare. Each time I have
> strong bands on the gel prior to cutting the bands, but everytime I
> purify and run the DNA on a gel to check there is nothing!!!
>
> Can anyone help?
> Thanks
> Hilary