Primer sequences:
Forward Primer (rpoCF):
5' - MAYGARAARMGNATGYTNCARGA - 3' Reverse Primer (rpoCR):
5' - GMCATYTGRTCNCCRTCRAA - 3' Illumina Sequencing Library Prep Protocol
PCR Master Mix
| Reagent | Per reaction |
|---|---|
| Platinum Hot Start MM | 10 µL |
| rpoCF (10 µM) | 0.5 µL |
| rpoCR (10 µM) | 0.5 µL |
| Molecular-grade H₂O | 10 µL |
| Template DNA | 4.0 µL |
| Total reaction volume | 25 µL |
Thermocycler Conditions
Lid Temperature: 105°C
Total Volume: 25 µL
| Temperature | Time | Cycles |
|---|---|---|
| 94°C | 3 min | |
| 94°C | 45 sec | |
| 39.5°C | 1 min | 40x |
| 72°C | 1:30 min | |
| 72°C | 10 min |