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Team:DTU-Denmark/Methods/PCR
From 2013.igem.org
(→Generel example on a normal PCR-program) |
(→Generel example on a normal PCR-program) |
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== Generel example on a normal PCR-program == | == Generel example on a normal PCR-program == | ||
# 98°C for 2:00 - Denaturing and warm up | # 98°C for 2:00 - Denaturing and warm up | ||
| - | # 98°C for 0:10 - Denaturing after each cycle | + | # 98°C for 0:10 or 0:20 - Denaturing after each cycle |
| - | # 59°C for 0:30 - Annealing | + | # 59°C for 0:30 to 1:00 - Annealing |
# 72°C for 2:00 - Elongation | # 72°C for 2:00 - Elongation | ||
# Go to number 2 - repeat 35 | # Go to number 2 - repeat 35 | ||
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Always use heated lid and set the machine to wait until the lid have the right temperature. If not default set to lid temperature to 104°C. | Always use heated lid and set the machine to wait until the lid have the right temperature. If not default set to lid temperature to 104°C. | ||
| - | The method will usually be changed with regard to annealing temperature and/or elongation temperature; depending the length of the PCR-product. Others methods can also be used such as | + | The method will usually be changed with regard to annealing temperature and/or elongation temperature; depending the length of the PCR-product. Others methods can also be used such as [https://2013.igem.org/Team:DTU-Denmark/Methods/PCR-ramp "ramp-PCR"], [https://2013.igem.org/Team:DTU-Denmark/Methods/PCR-touchdown "touchdown-PCR"],[https://2013.igem.org/Team:DTU-Denmark/Methods/PCR-touchUp "touchUp-PCR"], [https://2013.igem.org/Team:DTU-Denmark/Methods/PCR-gradient "gradient-PCR"] or a combination of these. In addition to these often used standard procedures there are tons of methods to do and improve your PCR and also for cloning and quantification purpose, e.g. Nested-PCR, Hot start-PCR, quantitative-PCR, Assembly-PCR and many more. |
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| + | {{:Team:DTU-Denmark/Templates/EndPage}} | ||
Latest revision as of 14:31, 29 August 2013
PCR
Generel example on a normal PCR-program
- 98°C for 2:00 - Denaturing and warm up
- 98°C for 0:10 or 0:20 - Denaturing after each cycle
- 59°C for 0:30 to 1:00 - Annealing
- 72°C for 2:00 - Elongation
- Go to number 2 - repeat 35
- 72°C for 5:00 - Final extension
- End
Always use heated lid and set the machine to wait until the lid have the right temperature. If not default set to lid temperature to 104°C. The method will usually be changed with regard to annealing temperature and/or elongation temperature; depending the length of the PCR-product. Others methods can also be used such as "ramp-PCR", "touchdown-PCR","touchUp-PCR", "gradient-PCR" or a combination of these. In addition to these often used standard procedures there are tons of methods to do and improve your PCR and also for cloning and quantification purpose, e.g. Nested-PCR, Hot start-PCR, quantitative-PCR, Assembly-PCR and many more.
