Haynes Lab:Notebook/Heathers Lab Notebook/2013/07/24

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Second Plasmid Procedure

  • Plasmid KA14 was selected.
  • Added 9μL of sterile H2O, following with 1μL plasmid (Note: The plasmid was almost empty, so the volume may have been less) to tube
  • Competent DH5-Alpha cells were obtained and thawed in an ice bath for ten minutes.
  • 50μL of these cells were then pipetted to the tube with plasmid and water for a total volume of around 60-70μL. The remaining cells were disposed.
  • Prewarmed agar plates were labeled with Amp resistance, the date 07/24/2013, initials HB, the strain and KA14. The total volume of cells were pipetted into agar plates using a measurement of 70μL.
  • Six sterile glass beads were used to spread the cells. The plates were then placed in the incubator at 37 degrees C for 12 hours.