Basic information
Date:
Wed, 2008-01-09 00:00
Template:
dna
Associated reaction(s):
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@5min, 40X(94ºC@30sec,52ºC@30sec, 72ºC@30sec)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 | DNA melting producing single-stranded DNA | 30 | 94 | |
Annealing | The primer binds to the template | 30 | 52 | |
Extension | The primer binds to the template | 30 | 72 | |
Final elongation | Any remaining single-stranded DNA is extended | 1 | 600 | 72 |
Hold | 1 | 8 |
Reversed primer(s):
People:
Products and Equipment:
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