Team:Glasgow/Notebook/Protocols

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Protocols

Edit This page shows our protocols for various lab procedures.

Creating Chemically Competent Cells

  1. Pipette 400μl of culture into 20ml of fresh broth
  2. Incubate the cultures in shaking 37⁰C water bath for 90 min
  3. Calcium chloride and centrifuge tubes must be cooled on ice.
  4. Pour culture into centrifuge tubes, return to ice.
  5. Centrifuge in a cold rotor for 2 min at 7000rpm (6000 G)
  6. Pour out the supernatant, being careful about the pellet
  7. Add 10μl of calcium chloride, immediately return to ice and leave for 1 hour (or longer)
  8. Repeat the centrifuge step, pour out supernatant
  9. Store on ice, add 1ml of calcium chloride. Resuspend and return to ice.

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