COI- LCO/HCO 20071105
Basic information
Date:
Mon, 2007-11-05 00:00
Gene:
Template:
dna
Procedure
PCR cocktail:
| Component | Volume (ul) | Mix (ul) |
| Forward primer | 1 | |
| Reversed primer | 1 | |
| Buffer | ||
| MgCl2 | ||
| dNTPs | ||
| Taq | ||
| DNA (if nested) | 1 | |
| ddH2O |
22 | |
| Total volume | 25 |
Forward primer(s):
Thermal cycling (abbreviated):
95ºC@5min, 40x(94ºC@15sec,46ºC@15sec, 72ºC@15sec),72ºC@10min
PCR machine:
GeneAmp
Thermal cycling:
| Step | Comment | Cycles | Time (sec) | Temperature (°C) |
| Initialization | For DNA polymerases that need hot-start heat activation | 1 | ||
| Denaturation | DNA melting producing single-stranded DNA | 1 | 300 | 95 |
| Amplification | 40 | |||
| Denaturation | 15 | 94 | ||
| Annealing | 15 | 46 | ||
| Extension | 15 | 72 | ||
| Final elongation | Any remaining single-stranded DNA is extended | 1 | 600 | 72 |
| Hold | 1 | 10 |
Reversed primer(s):
People:
Products and Equipment: