Quick and cheap +/- PCR screen?

dkim at unm.edu dkim at unm.edu
Wed Jun 8 12:46:00 EST 1994


<Query about quick assay of success of PCR reactions>

I read once, mayb e twice, papers in BioTechniques in which the success of 
the PCR was assessed by adding ethidium bromide to the reaction mixes. This 
can actually be done before the reaction is run, it does not inhibit the 
enzyme. Then, the tubes can be placed under a UV lamp to check for 
fluorescence. A further refinement was described in which the PCR was done in 
a 96-well format with fiber-optic monitoring of individual wells on a cycle-to-
cycle basis, so the rate of DNA synthesis could be assessed in all 96 wells 
at once.

Anyways, if you want to just know whether DNA is being made, this is one way
to go. There will be a little background fluorescence attributable to ssDNA
primers and the dNPTs (?), but that is much less than would be generated from
dsDNA products.

Daniel Kim   kim at flovax.lanl.gov



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