Team:Chiba/protocol/PCR

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>Protcol

PCR

  • Resuspend primer in Nuculease free water to 100 µM
  • PCR mix
  1. DNA template 1µL
  2. Fwd primer 10µL (final con. 10 pM)
  3. Rev primer 10µL
  4. 10x thermo pol buffer 10µL
  5. dNTP mix 10µL
  6. DNA pol. 1µL
  7. dH20 58µL


----------------------------
100µL

  • PCR cycle
  • Start: 94 °C for 5 min. (melt)
  • cycle: melt: 1 min.
  • anneal : 30 sec.
  • cycle end: extension:72 °C for 3.5 min.
  • 25 cycles
  • 72 °C for 10 min
  • End: keep at 6 °C forever
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