IGEM:Yale/2010/Protocols/Transformation

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Standard Transformation Protocol

  1. Preheat SOC broth at 37 °C
  2. Thaw competent cells on ice
  3. Aliquot 50-100 μL of cells into two pre-chilled 15 mL culture tubes (one for the control, one for the actual transformation)
  4. Add 1-2 μL of plasmid and swirl gently
  5. Incubate tubes on ice for 30 minutes
  6. Heat pulse tubes in 37°C degree water fro 30-40 seconds
  7. Incubate tubes on ice for 2 minutes
  8. Add 500 μL of preheated SOC broth to each tube and incubate for an hour at 37°C with shaking
  9. Spread 100 μL of each culture on a LB agar plate containing the appropriate antibiotic(s) and incubate overnight at 37°C
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