TAMRA as reporter in realtime-PCR

Bodychecker megacheck at hotmail.com
Wed Jul 11 14:31:08 EST 2001


anybody experience with low (or no) fluorescence signal in TAQMAN-PCR with
5' Tamra and 3' black hole quencher-2 labelled probes or other probes (like
molecular beacons) using TAMRA as reporter dye (NOT as quencher, as
frequently used)?

We have two TAQMAN probes that work fine when labelled with FAM as reporter.
Replacing this probe with the 5'TAMRA/3'blackholequencher-2 combination only
gives 'background signals' (nothing comparable to normal amplification
plots). We already titrated different concentrations of the probe, which did
not help.

I would be especially interested if this may be due to a low fluorescence of
the TAMRA dye in general.

THNX a lot in advance


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