Team:DTU-Denmark/PCR program design

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# Final extension for 10 min at 72⁰C
# 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)
# Cooling down to 4⁰C (usually time is set as eternity so it can be stored in the machine until you can take it out)
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Revision as of 14:48, 18 September 2011

Basic PCR program design

Basic PCR program design

  1. Initial denaturation for 2 minutes at 95⁰C
  2. denature for 1 minute at 95⁰C
  3. Anneal primers for 30 seconds at temperature ~5⁰C below melting temperature of primers.
  4. 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.
  5. Repeat steps 2-4 for 25-30 cycles.
  6. Final extension for 10 min at 72⁰C
  7. Cooling down to 4⁰C (usually time is set as eternity so it can be stored in the machine until you can take it out)

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