Silver: Bacterial Transformation

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  1. Thaw competent cells (TOP10 from Invitrogen) on ice.
  2. Place 10-15 µL cells in pre-chilled Eppendorf tubes.
  3. Add 2 µL BioBrick vector (either native or ligated; so that vector is ~10-20% of the final volume), and chill on ice for 30 min.
    • Do not pipet or vortex.
  4. Heat shock at 42 °C for 30 s.
  5. Incubate on ice for 2 min.
  6. Add 170 µL SOC medium (~10 x volume; Invitrogen), and shake at 37 °C for 15 min for Amp selection, or 30 min for Kan selection.
  7. Add all media to a selectable marker LB plate. Use glass beads to streak the plate.
  8. Incubate at 37 °C. Transformants should appear within 12 hrs.