cleaning PCR primers
Biochemistry Lab computer
biochem at vcp.monash.edu.au
Sun Dec 11 19:03:23 EST 1994
We have just made some oligos for PCR on a 50pmol scale, cleaved them from
the column, deprotected and dried them down. When they are redissolved in
water there is visible rubbish in the tube, which does spin down. I have
tried using the clear supernatant, diluted, in a PCR reaction and get
nothing. My question is, does anyone know if dirty primers can inhibit the
PCR reaction? And if so what is the best method (and simplest) for
cleaning them up? I have, in the past, purified oligos off an acrylamide
gel, but when I was using a larger scale synthesis. My concern is, that
there may not be enough oligo to purify off a gel, considering the losses. I
have previously not needed to purify the 0.2nmol scale synthesis, finding
they always worked.
Thanks in advance.
Jo Caine
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