Team:UC-Santa Cruz-BioE/Notebook

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Notebook

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Important Protocols

Glycerol Stock Preparation

  1. Inoculated TSB with MR-1 (either from plate or glycerol stock) and grow overnight.
  2. add glycerol to culture to final concentration of 80% glycerol.
  3. place 1ml in 1.5ml freezer safe eppendorf tubes (do not fill eppendorfs with more than 1.5ml to allow for expansion by freezing).

Microbial Fuel Cell Assembly

  • cut two 3 inch PVC tubes .
  • sandwich them with nafion cation exchange membrane between them.
  • add caulking around both sides of membrane-tube junction.
  • let dry for 15 minutes.
  • repeat step 3 and 4.
  • cut off excess membrane around junction.
  • add one more layer to seal up junction.
  • let dry overnight.
  • cut 2.5in-3in x 12in of carbon cloth
  • roll around tip of finger.
  • weave titanium wire close to end edge of roll.
  • fold titanium wire up the length of the roll.
  • twist excess wire around itself.
  • cut wire at .5in-1in.
  • push through stopper.
  • seal hole from wire with caulking.
  • let dry overnight. (two of these make up the cathode and the anode)
  • pour potassium ferricyanide in one chamber of the cell.
  • carefully push stopper with carbon cloth to plug the chamber.
  • push hollow needle through stopper to let air out of chamber.
  • pour overnight culture of MR-1 into other chamber.
  • place glass slide in chamber.
  • carefully push stopper with carbon cloth to plug the chamber.
  • push new sterile hollow needle in stopper into air bubble in chamber to let out any CO2 bacteria will produce.
  • open ExcelINX up on computer.
  • connect electrodes from DMM to cathode and anode (remember which channels are being used).
  • start ExcelINX.

Plasmid Prep