Probe labelling by PCR

[Dan Pereira]

Thomas Urbig <Thomas at hastingslab.harvard.edu> wrote:
>
> Has onybody experience labeling gene probes by PCR and can provide 
> reaction conditions for this. It should also be possible to make single 
> stranded probes. If so, do you use a single stranded PCR product as 
> template or is a standard (double strand) PCR product as template fine. 
> Is the lower concentration of the 32P nucleotide a problem or do I have 
> to add cold nucleotide? 
> Any advice is welcome, thanks in advance
>   
>                Thomas.
> 
> 
I have had success with 5'-end labeled oligo primers and inclusion of alpha 
labeles nucleotide in the reaction. If however, you choose to include 
alpha-nucleotide in the reaction you should supplement with at least
20 uM cold nucleotide.  Best of Luck.
Dan