IGEM:PennState/2006/Transformation

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Contents

Transformation

Media/Reagents

  • Competent cells
  • SOC1
  • Antibiotic LB plates

Pre

  • Add plasmid DNA to alloquats from electrocompetent cell prep
  • Transfer DNA and Cells into electroporator cuvettes

Electroporation

  1. Shock Cells, monitor discharge time
  2. Add SOC to cuvettes (~400)
  3. Transfer to culture tubes, shake at 37 for 1 hour (depending on antibiotic marker)
  4. Plate ~200uL of SOC/Cells onto agar plate

1To make SOC, add 20 μL 500 mM filter-sterilized glucose to 500 uL SOB (20 mM final glucose concentration)

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