Table 1.
Optimized conditions for prfA qPCR.
| Polymerase | Initial denaturation [°C – min] |
Denaturation [°C – min] |
Annealing/ Annealing-Elongation [°C – min] |
Elongation [°C – min] |
Cycles | MgCl2 [mM] |
|---|---|---|---|---|---|---|
| prfA-standard-conditions | 94 - 2.00 | 94 - 0.15 | 64 - 1.00 | – | 45 | |
| Platinum™Taq DNA Polymerase | 94 - 2.00 | 94 - 0.15 | 64 - 1.00 | – | 45 | 3.5 |
| TaqMan™ Fast Advanced Master Mix | 95 - 5.00 | 95 - 0.30 | 57 - 0.30 | 72 - 1.00 | 50 | (buffer) |
| AmpliTaq Gold™ DNA Polymerase | 95 - 10.00 | 94 - 1.00 | 59 - 2.00 | – | 50 | 1.5 (buffer) |
| Maxima Hot Start PCR Master Mix | 95 - 5.00 | 95 - 0.30 | 57 - 0.30 | 72 - 1.00 | 50 | 2 (buffer) |
| SsoAdvanced™ Universal Probes Supermix | 94 - 2.00 (95 - 10.00) |
94 - 0.15 (94 - 1.00) |
64 - 1.00 (59 - 2.00) |
– (-) |
45 (50) |
(buffer) |
| Maxima Probe/ROX qPCR Master Mix | 95 - 7.00 | 95 - 0.30 | 55 - 0.30 | 72 - 1.30 | 50 | (buffer) |
| iTaq™ Universal Probes Supermix | 94 - 2.00 (95 - 10.00) |
94 - 0.15 (94 - 1.00) |
64 - 1.00 (59 - 2.00) |
– (-) |
45 (50) |
(buffer) |
| Hot Start Taq DNA Polymerase | 95 - 2.00 | 95 - 0.15 | 51 - 0.30 | 68 - 1.00 | 50 | 4.5 |
| BioThermAB™ Hot Start Taq DNA Polymerase | 95 - 3.00 | 95 - 0.30 | 57 - 0.30 | 72 - 1.00 | 50 | 3.5 |
| BioThermPlus™ DNA Polymerase | 95 - 2.00 | 95 - 0.20 | 51 - 0.20 | 72 - 0.30 | 50 | 2.5 |
| BioThermBio™ Taq DNA Polymerase | 95 - 2.00 | 95 - 0.20 | 51 - 0.20 | 72 - 0.30 | 50 | 2.5 |