7. Real-time PCR Flashcards
1
Q
DNA probe - what’s on the 5’ end and 3’ end?
A
5’ - reporter
3’ - quencher
2
Q
Why can’t melt-curve analysis be done on TaqMan probes?
A
TaqMan probes are degraded for fluorescence detection (chewed up by Taq)
No probes to melt off of amplicons = can’t get Tm info
3
Q
In terms of real-time PCR, what is ‘threshold’
A
Establishes the point where measurements are made - therefore placed somewhere in exponential phase of all amplification curves
4
Q
Relationship between Ct (cycle threshold) and number of starting target copies in real-time PCR
A
Inversely proportional
Higher Ct = lower starting target copy
