PCR reamplification problem

Rosemary Brown 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. 
Rose Brown
Massey University 
New Zealand



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