Non-radioactive PCR

Daniel Kim dkim at nmsu.edu
Fri Aug 11 14:12:07 EST 1995


In article <9508102038.AA18031 at intnet.upj.com> rvdeshp0 at INTNET.UPJ.COM writes:
>I would like to qualitatively determine the positivity or negativity of a
>sample for PCR with a given set of primers.  I plan to use biotinylated or
>fluorescinated primers (or a combination of both) for PCR, and detect the
>final product in an ELISA using streptavidin-coated plate and
>anti-fluorescin-AP conjugate.  There are a few papers in the journal -
>Biotechniques, that describe parallel procedures.  Does anyone have any
>suggestions on the "do's and don'ts" of such techniques ?  Also, it would be
>of great help to know how to avoid picking up signal from false-positive
>PCR by-products ?  Any help will be greatly appreciated.  Thanks.
>


Hello.
If all you want is a +/- answer, you might want to consider adding 
ethidium bromide to your PCR reaction mixes (no other label needed).  
After the run, just look for UV-fluorescence, indicating the presence of 
dsDNA products.  (have I said this before?)

Daniel Kim




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