components | SYBR® Premix Ex Taq™ II | forward and reverse primer (10 μM) | template DNA | sterile deionized water | total |
---|---|---|---|---|---|
volume | 12.5 μl | 2 μl | 1 μl | 9.5 μl | 25 μl |
reaction procedures | predenaturation | 40Â cycles | extension | ||
 |  | denaturation | annealing | extension |  |
Temperature (°C) | 95 | 94 | optimal temperature | 72 | 72 |
Time (s) | 60 | 15 | 30 | 30 | 10 |