| Component | Amount |
| 2× Reaction buffer | 25 μL |
| 5hmC dNTP Mix (10 mM each dNTP) | 1 μL |
| 5hmC control primer-F (10 μM) | 2 μL |
| 5hmC control primer-R (10 μM) | 2 μL |
| pUC 19 vector (1 ng/–L) | 1 μL |
| ZymoTaq™ DNA Polymerase (5 U/μL) | 0.4 μL |
| Milli-Q water | 18.6 μL |
| Final volume | 50 μL |