cleaning PCR primers,correction

Jo Caine 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:
>Path: harbinger.cc.monash.edu.au!biochem.vcp.monash.edu.au!biochem
>From: biochem at vcp.monash.edu.au (Biochemistry Lab computer)
>Newsgroups: bionet.molbio.methds-reagnts
>Subject: cleaning PCR primers
>Date: Mon, 12 Dec 1994 00:03:23 GMT
>Organization: Victorian College of Pharmacy
>Lines: 13
>Message-ID: <biochem.43.2EEB934A at vcp.monash.edu.au>
>NNTP-Posting-Host: biochem.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. 
>Jo Caine

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

Thanks again.
Jo Caine



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