PCR-PROPDUCT PURIFICATION

Carlisle Landel landel at helios
Tue Nov 22 10:50:40 EST 1994


In article <3apv03$o2s at lyra.csx.cam.ac.uk>,
Charlie Wright (Genetics) <cw117 at mole.bio.cam.ac.uk> wrote:
>mbjjb at s-crim1.dl.ac.uk (J.J. Baker) writes:
>
>>sl6dp at cc.usu.edu wrote:
>>: Going through the literature I have found that it is possible to ream-
>>: plify PCR products directly from the band after selection on a low-melting
>>: agarose gel. I am not sure if I understand it: it is really possible
>>: to run the PCR with a portion af the agarose band, without even
>>: taking the DNA out????. Anyaway, which is a good method for PCR product
snip
>
>>Yes it is! Though I can't remember the exact referencefor this technique,
>>this is the method I use; I pierce the bands of the gel with a wide bore
>>needle and then put the needle into the reaction buffer for PCR then leave for 
>>about ten minutes or the length of your tea break at room temperature
>>Then simply take out the needles, add your Taq and run for 5 more cycles
>>than usual.
>
snip
>A chap in our lab also uses this technique.  I believe he actually blows 
>the plug of agarose (low melt) into the eppendorf tube and then adds the 
>reagents for PCR.  You'll find that the lmp agarose will melt quite 
>readily at 96c and then it will be too dilute to cause a problem, usually.
>

Actually, you can just poke the band with a toothpick, swirl it around
in your PCR reaction mix, and go.

Carlisle Landel



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