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Team:DTU-Denmark/PCR program design
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== Basic PCR program design == | == Basic PCR program design == | ||
# Initial denaturation for 2 minutes at 95⁰C | # Initial denaturation for 2 minutes at 95⁰C | ||
Revision as of 14:48, 18 September 2011
Basic PCR program design
Basic PCR program design
- Initial denaturation for 2 minutes at 95⁰C
- denature for 1 minute at 95⁰C
- Anneal primers for 30 seconds at temperature ~5⁰C below melting temperature of primers.
- Extend DNA at 72⁰C using each 1-2 minutes per kilobase of product, depending on whether you are using a polymerase with proofreading capabilities. If in doubt see manufacturers instructions.
- Repeat steps 2-4 for 25-30 cycles.
- Final extension for 10 min at 72⁰C
- Cooling down to 4⁰C (usually time is set as eternity so it can be stored in the machine until you can take it out)
