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Pf0-1 Pseudomonas Fluorescens Electrocompetency Prep

  • Innoculate 6 mL of liquid LB or Pseudomonas Minimal Media with Pf0-1 and grow overnight
  • Split overnight into 1.5 mL aliquots and centrifuge at 16,000x g, discard supernatant
  • Resuspend cell pellet with 1 mL of 300 mM sucrose in water solution
  • Centrifuge at 16,000x g and discard supernatant
  • Repeat resuspension and centrifuge step
  • Resuspend cell pellet with 50 uL or 100 uL 300 mM sucrose in water solution

Pf0-1 Pseudomonas Fluorescens Electroporation

  • Retrieve a 50 uL or 100 uL vial of electrocompentence prepped cells
  • Add 2 uL of DNA directly into vial of cells and mix by flicking
  • Transfer mixture to a 2 mm Electroporation cuvette
  • On BioRad Gene Pulser, pulse with 2.5 kV, 200 Ohm, 25 uF settings
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