PCR reamplification problem
Rosemary.Brown.3 at uni.massey.ac.nz
Mon Jun 23 13:05:30 EST 1997
In response to Allen Sylvestor's comments on the reamplification of PCR
products using gel stabs..
> Disposable plastic pipette tips would probably
> work about as well, except if the plug moves up away from the opening, it
> is a hassle to get it out. Similarly, when I tried to suck the plug up
> into the pipette, it went too far and would not come out without breaking
> the pipette.
I use a similar method of reamplification from Nusieve agarose using
disposable plastic pipette tips, for a 1 kb product. Just invert them
and centrifuge for a minute or so to get the plug out.No worries. I have
used the gel plug directly in a PCR reaction and got products OK for
sequencing, except its a tad viscous. I have had success using a sterile
unwound , metal paper clip, stabbed into the gel and dunked into the PCR
mixture for a bit.
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