Basic information
Procedure
PCR cocktail:
Component | Volume (ul) | Mix (ul) |
Forward primer | ||
Reversed primer | ||
Buffer | ||
MgCl2 | ||
dNTPs | ||
Taq | ||
DNA (if nested) | ||
ddH2O |
||
Total volume |
Forward primer(s):
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 | ||
Amplification | Z | - | - | |
Denaturation | DNA melting producing single-stranded DNA |
- | ||
Annealing | The primer binds to the template | - | ||
Extension | The DNA polymerase synthesizes new DNA | - | ||
Final elongation | Any remaining single-stranded DNA is extended | 1 | ||
Final hold | Short-term storage | 1 | - |
Notes:
20 ul PCR-product was Gel Extracted and eluted in 35ul EB according to the QIAquick Gel Extraction Kit spin protocol.
Reversed primer(s):
File attachments:
People:
Products and Equipment:
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