Jones Lab:Protocols:PCR

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PCR

Add the following to a PCR tube (Master mix if necessary)

  1. 10 μL H2O
  2. 2 μL 10x PCR buffer
  3. 2 μL 2.5mM dNTPs (each)
  4. 2 μL 20mM MgCl2
  5. 1 μL Forward primer (10μM)
  6. 1 μL Reverse primer (10μM)
  7. 1 μL taq polymerase
  8. 1 μL DNA

Mix gently, then setup PCR cycles

  1. 94°C for 2 mins
  2. 94°C for 30 secs
  3. 58°C for 15 secs
  4. 72°C for 1 min per Kb of amplicon
  5. Go to #2 35 times
  6. 72°C for 5 mins
  7. 10°C hold
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