Non-radioactive PCR

Torsten Boerchers borcher at uni-muenster.de
Wed Aug 16 04:46:55 EST 1995


>dkim at nmsu.edu (Daniel Kim) wrote:
>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
>

A similar approach for "Direct selection of positive recombinant
clones by PCR" was published by S. Pampfer in BioTechniques 
15, 590-592(1993) i.e., the nov/dec Edition of 
Biotechniques Europe (for the many subscribers of this nice 
and free! journal).

Torsten
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