Table 3 Sequencing reaction
Reagent | Reaction volume | |
BigDye Terminator Ready Reaction Mix | 4 μl | |
Template (100 ng/ul) | 1 μl | |
Primer (10 pmol/λ) | 2 μl | |
Milli-Q water | 3 μl | |
PCR condition for sequencing | ||
Step | Time and temp | Cycles |
Initial denaturation | 96 °C for 5 min | 25 cycles |
Denaturation | 96 °C for 30 s | |
Hybridization | 50 °C for 30 s | |
Elongation | 60 °C for 1.30 min | |