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Team:DTU-Denmark/PCR program design
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(Created page with "== PCR program design == Step 1: Initial denaturation for 2 minutes at 95⁰C Step 2: denature for 1 minute at 95⁰C Step 3: Anneal primers for 30 seconds at temperature ~5⁰C ...")
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(Created page with "== PCR program design == Step 1: Initial denaturation for 2 minutes at 95⁰C Step 2: denature for 1 minute at 95⁰C Step 3: Anneal primers for 30 seconds at temperature ~5⁰C ...")
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Revision as of 14:41, 18 September 2011
PCR program design
Step 1: Initial denaturation for 2 minutes at 95⁰C Step 2: denature for 1 minute at 95⁰C Step 3: Anneal primers for 30 seconds at temperature ~5⁰C below melting temperature of primers. Step 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. Step 5: Repeat steps 2-4 for 25-30 cycles. Step 6: Final extension for 10 min at 72⁰C Step 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)
