A-tailing of PCR products

[Simon Dawson]

cfb7 at psu.edu wrote:
> 
> Hi,
> I am using a degenerate PCR strategy to amplify fragments of expressed
> members of a subfamily of genes; I am ligating the products into a
> T-vector.  I use a 15' 72C incubation at the end of the PCR cycles to
> add the A-tails.  Is this sufficient to add A-tails to my products or
> should I purify the products and A-tail them in a subsequent reaction
> in which dATP is the only nucleotide available?  Thanks, C-
> 
> Christie Blackman          cfb7 at psu.edu

	I routinely use a 15' @ 72C step and this adds the As fine in a Taq-based PCR reaction.

-- 

Dr. Simon Dawson       TEL:+44 (0)115 9249924 ex. 44787
Dept. of Biochemistry  FAX:+44 (0)115 9422225
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