cleaning PCR primers,correction

Steve Hopkins shopkins at fs1.ho.man.ac.uk
Mon Dec 12 17:38:43 EST 1994


In article <JO_C.44.2EEBD151 at vcp.monash.edu.au> JO_C at vcp.monash.edu.au (Jo Caine) writes:
>From: JO_C at vcp.monash.edu.au (Jo Caine)
>Subject: cleaning PCR primers,correction
>Date: Mon, 12 Dec 1994 04:28:02 GMT

>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? >>Jo Caine

>Sorry, when I posted this message the terminal put in a bogus email address.
>Jo Caine

Repeat of the message I posted on your 'bogus' address.

We received some commercially prepared primers in a similar state, which did 
not work properly. The issue was not resolved entirely, but seemed to be 
due to material coming off the column. The material from the column 
resulted in a spuriously high OD value, resulting in over dilution of the 
primers (i.e. it was difficult to know how much primer we had), rather than 
the dirty primers interfering with PCR. 








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