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Team:DTU-Denmark/Methods/PCR-ramp
From 2013.igem.org
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Ramp-PCR is when you make the PCR-program to ramp the temperature down from x to y so you will get a specific annealing. For instance: | Ramp-PCR is when you make the PCR-program to ramp the temperature down from x to y so you will get a specific annealing. For instance: | ||
| + | # 98°C for 2:00 - Denaturing and warm up | ||
| + | # 98°C for 0:10 - Denaturing after each cycle | ||
| + | # 65°C for 0:05 - Annealing | ||
| + | #* ramp 0.1°C/s until 60°C | ||
| + | # 72°C for 2:00 - Elongation | ||
| + | # Go to number 2 - repeat 35 | ||
| + | # 72°C for 5:00 - Final extension | ||
| + | # End | ||
| + | |||
| + | A program like that will typically help you if your primers have annealing temperatures in the area between 65°C and 60°C but you don't know the best single step temperature. | ||
Revision as of 18:38, 26 June 2013
Ramp-PCR
Ramp-PCR is when you make the PCR-program to ramp the temperature down from x to y so you will get a specific annealing. For instance:
- 98°C for 2:00 - Denaturing and warm up
- 98°C for 0:10 - Denaturing after each cycle
- 65°C for 0:05 - Annealing
- ramp 0.1°C/s until 60°C
- 72°C for 2:00 - Elongation
- Go to number 2 - repeat 35
- 72°C for 5:00 - Final extension
- End
A program like that will typically help you if your primers have annealing temperatures in the area between 65°C and 60°C but you don't know the best single step temperature.
