Basic information
Procedure
PCR cocktail:
| Component | Volume (ul) | Mix (ul) |
| Forward primer | 0,5 | |
| Reversed primer | 0,5 | |
| Buffer | ||
| MgCl2 | ||
| dNTPs | ||
| Taq | ||
| DNA (if nested) | 2 | |
| ddH2O |
22 | |
| Total volume | 25 |
Forward primer(s):
Thermal cycling (abbreviated):
95ºC@4min, 35x(94ºC@30sec, 50ºC@30sec, 72ºC@30sec), 72ºC@10min
Thermal cycling:
| 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 | 1 | ||
| 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 | - | 12 |
PCR program:
COI
Reversed primer(s):
File attachments:
People:
Products and Equipment:
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