The polymerase chain reaction

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Presentation transcript:

The polymerase chain reaction CHAPTER TWENTY-THREE The polymerase chain reaction

Figure 23.01

Figure 23.04

Figure 23.05

Figure 23.02

Value for indicated DNA polymerase Property Value for indicated DNA polymerase KOD DNA polymerase Pfu DNA polymerase Taq DNA polymerase Origin Archaea Bacteria Deduced molecular mass (kDa) 90.0 90.1 93.9 Optimum temperature (ºC) 75 Optimum pH at 75ºC 6.5 8.0-8.5 Thermostability (half-life) 95ºC,12 h; 100ºC,3.0 h 95ºC, 6h; 100ºC, 2.9h 95ºC, 1.6 h 5'→3' exonuclease activity - + 3'→5' exonuclease activity Terminal transferase activity Processivity (bases) >300 <20 ND Elongation rate (bases/s) 106-138 25 61

Reverse transcription PCR

Figure 23.15

Figure 23.16

Figure 23.08

PCR for cloning

One-step assembly PCR fusion PCR

PCR for detection

Real-time fluorescent PCR

Figure 23.25 http://www.premierbiosoft.com/tech_notes/molecular_beacons.html