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DNA probe - what's on the 5' end and 3' end?

5' - reporter
3' - quencher


Why can't melt-curve analysis be done on TaqMan probes?

TaqMan probes are degraded for fluorescence detection (chewed up by Taq)

No probes to melt off of amplicons = can't get Tm info


In terms of real-time PCR, what is 'threshold'

Establishes the point where measurements are made - therefore placed somewhere in exponential phase of all amplification curves


Relationship between Ct (cycle threshold) and number of starting target copies in real-time PCR

Inversely proportional

Higher Ct = lower starting target copy