Table 2 PCR reaction components.
| Reaction 1 | Reaction 2 | |
|---|---|---|
| Template plasmid DNA | ~500 ng | ~500 ng |
| Forward primer | 40 pmol | - |
| Reverse primer | - | 40 pmol |
| MgCl2 | 0.2 mM | 0.2 mM |
| dNTPs | 0.2 mM | 0.2 mM |
| Pwo Master DNA polymerase | 1.25 U | 1.25 U |
| Tris buffer, pH 7.5 | 10 mM | 10 mM |
| Final volume | 25 μl | 25 μl |