How to add an A with Taq
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.
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