Difference between revisions of "Team:Paris Bettencourt/Protocols/PCR purification"

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Latest revision as of 18:14, 14 August 2015

    PCR purification protocol
  • Add 5 volumes of resuspension buffer to 1 volume of PCR product in an 1.5mL microcentrifuge tube, mix by pipetting up and down
  • Transfer in a centrifugation column
  • Centrifuge 1 min at 14000 rpm
  • Throw the filtration product
  • Add 700μL of washing solution
  • Centrifuge 1 min at 14000 rpm
  • Throw the filtration product
  • Add 500μL of washing solution
  • Centrifuge 1 min at 14000 rpm
  • Throw the filtration product
  • Centrifuge 1 min at 14000 rpm
  • Throw the filtration product
  • Put the column in a sterile 1.5mL microcentrifuge tube
  • Add 45μL of DNAse/RNAse free water on the membrane
  • Wait 2 minutes
  • Centrifuge 2min at 10000rpm
  • Discard the column, DNA is saved in water </td>