PCR COI- LCO/HCO 20071022
Basic information
Date:
Sun, 2007-10-21 23:00
Gene:
Template:
dna
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):
94ºC@5min, 40x(94ºC@15sec, 46ºC@15sec, 72ºC@15sec), 72ºC@10min
Thermal cycling:
Step | Comment | Cycles | Time (sec) | Temperature (°C) |
Initialization | For DNA polymerases that need hot-start heat activation | 1 | 300 | 94 |
Denaturation | DNA melting producing single-stranded DNA | 1 | ||
Amplification | 40 | - | - | |
Denaturation | DNA melting producing single-stranded DNA |
- | 15 | 94 |
Annealing | The primer binds to the template | - | 15 | 46 |
Extension | The DNA polymerase synthesizes new DNA | - | 15 | 72 |
Final elongation | Any remaining single-stranded DNA is extended | 1 | 600 |
72 |
Final hold | Short-term storage | 1 | - | 12 |
Reversed primer(s):
People:
Products and Equipment: