which PCR machine?
ludla at aol.com
Thu Oct 9 05:55:34 EST 1997
If you are considering the PE "Taq-Man" assay, be advised that you must have
specifically designed probes for the system. This means you must know, in
advance, the sequence of your target, design the probes which will require
both a fluorophor and quencher molecule on the probe.
The assay works because the quencher proximity to the fluorphor prevents
(within reason) fluorescence UNTIL the Taq digests the probe off the target as
it extends the new strand.
Needless to say, as with all things, there are drawbacks: background must be
accounted for, probe costs can be significant, co-optimization of probe
binding and primer binding must be determined, etc.
This is not meant to discourage, but to enlighten! Remember always that
Marketing can make it sound EASY AS FALLING OFF A LOG!
More information about the Methods