hot PCR

[Rob Jordan]

In article <330CA1D1.20C9 at niehs.nih.gov>, NIEHS, Mail, Drop, 7-10, Research, Triangle, Park, NC, 27709 wrote:
>I need a protocol for generating a radioactive probe by PCR 
>(concentrations of hot and cold nucleotides, particularly).  
>THanks, Caroline

Check out:

Mertz and Rashtchian,  Anal. Biochem., 221, p160-165, 1994

If I recall conrrectly using a cold dNTP concentration about twice that of the 
hot dNTP concentration gave the best amplification and labeling.  The trick is 
that the hot dNTP concentration from places like Amersham or DuPont is really 
low.  

Rob
rjordan at u.washington.edu