How to add an A with Taq

Robert Edwards redwards at pobox.upenn.edu
Fri May 12 17:57:32 EST 1995


Juan Pablo Labrador <Labrador at embl-heidelberg.de> wrote: 
>Does anyone have a reliable protocol to add terminal As to PCR products 
>produced Pfu polimerase or any other than Taq. 
>I think that it is possible to do this incubating the PCR product for  
>some time at 72 degrees with Taq polimerase. 
>What I don't know is how much time should I incubate 
>if I have to add dATP and the final concentration 
>If I have to purify the PCR product before adding the As with the Taq  
>or I can use the Pfu polimerase mix without any further treatment. 
> 
>Thanks in advance for your comments.  
> 
 
Apparently a 15 minute incubation time after the final extension, without 
purifying or adding extra dATP is sufficient. I haven't tried this though 
 
Try calling your supplier, and asking them. 
 
Rob




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