small fragment of DNA PCR Product elution kit difficulty

[Jose de las Heras]

"V Arunachalam" <vadivelarunachalam at yahoo.com> wrote in message 
news:mailman.1063.1158411969.20007.methods at net.bio.net...
> Hi,
>  I am facing problem while elutung a PCR product amplified from genomic 
> DNA of 200 bp size fragment. The DNA recovery is low. Could anyone suggest 
> me good kit / modification in the protocol for getting high recovery of 
> DNA.


Try digestion of the agarose by enzymes such as Agarase/AgarAce etc 
(different names for different companies). That always worked very well for 
me.

A simple electroelution may work too (just put the cut band into a small 
piece of dialysis tubing with a little buffer, and run it for a few minutes 
in teh tank... recover the DNA in the buffr by precipitation).

Lately I have been needing to purify and clone many PCR fragments, and I got 
tired of the columns and their poor yield. I simply freeze the bands for 
15-20 minutes, then thaw them and spin them in an eppendorf at max speed for 
a couple of minutes. Use the "juice" for ligations, it works very well.

I hope any of these help.

Jose
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