18S S30/5FK 20110510
Component | Volume (ul) | Mix (ul) |
Forward primer | 0.5 | |
Reversed primer | 0.5 | |
Buffer | ||
MgCl2 | ||
dNTPs | ||
Taq | ||
DNA | 2 | |
ddH2O |
22 | |
Total volume | 25 |
Step | Comment | Cycles | Time (sec) | Temperature (°C) |
Initialization | For DNA polymerases that need hot-start heat activation | 1 | 240 | 95 |
Denaturation | DNA melting producing single-stranded DNA | |||
Amplification | 35 | - | - | |
Denaturation | DNA melting producing single-stranded DNA |
- | 30 | 94 |
Annealing | The primer binds to the template | - | 30 | 50 |
Extension | The DNA polymerase synthesizes new DNA | - | 30 | 72 |
Final elongation | Any remaining single-stranded DNA is extended | 1 | 600 | 72 |
Final hold | Short-term storage | 1 | - | 4 |