Table 2.
The reaction system and procedure of droplet digital PCR
| Component | Volume Per Reaction, μL | Final Concentration | Cycling Step | Temperature, °C | Time | Number of Cycles | Other Reaction Conditions |
|---|---|---|---|---|---|---|---|
| 2 × QX200 ddPCR EvaGreen Supermix | 10 | 1× | Enzyme activation | 95 | 5 min | 1 | Ramp rate: 2 °C/s Lid temperature: 105 °C Set the sample volume to 40 μL |
| Forward primer (2 μM) | 1 | 100 nM | Denaturation | 95 | 30 s | 40 | |
| Reverse primer (2 μM) | 1 | 100 nM | Annealing and extension | 60 | 1 min | 40 | |
| Diluted cDNA template | 2 | 100 ng/μL | Signal stabilization | 4 | 5 min | 1 | |
| DNase-free water | 6 | — | 90 | 5 min | 1 | ||
| Total volume | 20 | — | Hold | 4 | Infinity | 1 |