Probe labelling by PCR
mgs6145 at student.health.ufl.edu
Thu Nov 30 17:18:39 EST 1995
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
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.
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