Table 4.
Composition of reaction master mix for Platinum Taq DNA Polymerase High Fidelity
| Component | Volume in Each Reactiona |
|---|---|
| 10X High Fidelity PCR buffer | 5 µl |
| MgSO4 (50 mM) | Variable (see Table 2) |
| dNTP (10 mM each) | 1 µl |
| Sense primer (10 µM) (see Table 2) | 2 µl |
| Antisense primer (10 µM) (see Table 2) | 2 µl |
| DMSO or 5 M betaine solution | Variable (see Table 2) |
| Platinum Taq DNA Polymerase High Fidelity (5 U/µl) | 0.5 µl |
| Template DNA | Added in later step in protocol |
| Water | Bring final volume including DNA to 50 µl |
a
The volume for a single reaction is 50 µl so multiple the number of amplification reactions desired by 50 to determine how much reaction master mix to make. Always make more than you need to account for losses during pipetting.