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| Description: The TaqMan approach to quantitative real-time PCR (qPCR) involves a pair of PCR primers along with a probe sequence complementary to the target gene. , Primer 6‘ f 5 — 3: 3’ 5' 5’ 3’ Primer 1. Hybridization. Forward and reverse PCFl primers bind to denatured target DNA. TaqMan probe with reporter (Fl) and quencher (O) dye binds to target DNA between the primers. When probe is intact, emission by the reporter dye is quenched. 5’ 3' 3’ 5' 5’ —F 2' DNA polymerase 2. Extension. As DNA polymerase extends the forward primer, it reaches the TaqMan probe and cleaves the reporter dye from the probe. Released from the quencher, the reporter can now emit light excited by a laser. 0 0 C . . 3 ngh gene expresswn ‘45 ‘6 E Low gene expression 0 l l l l l l l l l 4 8 12 16 20 24 28 32 38 40 Cycles 3. Detection. Emitted light from the reporter is detected and interpreted to produce a plot that quantitates the amount of PCR product produced with each cycle. Picture Stats: Views: 199 Filesize: 176.06kB Height: 1056 Width: 940 Source: https://biology-forums.com/index.php?action=gallery;sa=view;id=44425 |