cleaning PCR primers,correction
JO_C at vcp.monash.edu.au
Sun Dec 11 23:28:02 EST 1994
In article <biochem.43.2EEB934A at vcp.monash.edu.au> biochem at vcp.monash.edu.au (Biochemistry Lab computer) writes:
>From: biochem at vcp.monash.edu.au (Biochemistry Lab computer)
>Subject: cleaning PCR primers
>Date: Mon, 12 Dec 1994 00:03:23 GMT
>Organization: Victorian College of Pharmacy
>Message-ID: <biochem.43.2EEB934A at vcp.monash.edu.au>
>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.
Sorry, when I posted this message the terminal put in a bogus email address.
The real one is as follows.
JO_C at VCP.MONASH.EDU.AU
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