Team:Grinnell/Notebook/Protocols

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===Competent Cells===

  1. Inoculate 500mL LB with 2mL overnight culture. Incubate with shaking to early log phase (~5x10^8cells/mL, OD600 = 0.3-0.5).
  2. Chill cells on ice for 15-120min.
  3. Pellet cells in a prechilled sterile centrifuge tube by centrifugation at 5-8krpm for 5min at 4 degrees C. Discard supernatant.
  4. Completely resuspend cells in 20mL cold 100mM CaCl2 (10% glycerol), and incubate on ice for 3hr.
  5. Harvest cells by cetrifugation. Discard supernatant.
  6. Gently resuspend cells in 0.5mL cold 100mL CaCl2 (10% glycerol). Incubate on ice for at least 1hr and freeze at -80 degrees C.

Plasmid

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