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Protocol
PCR-protocol
Acc.no:
1VEPSEX
|
Published:
2010-03-22
by
jennie
Keywords: PCR
PCR reactions are individually designed depending on the primers Tm and the specific Taq enzyme used.
Instructions
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A typical protocol will comprise:
- A denaturation step at 95 C.
- An annealing step between 50-70 C, depending on the primer.
- A final polymerization step at 72 C with a length depending on the fragment to amplify.
These three steps are looped in a number of cycles from 25 to 35 or more depending on the template.
The concentration of the different reagents (dNTP, primers, buffer, water, template) is recommended by the manufacturers.
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Example PCR-protocol
Step °C Duration [min] 1. 95 10 2. 95 1 3. 66 1 4. 72 2.30 5. Go to 2 25 times 6. 95 1 7. 72 7 8. 15 for ever
