Team:UC-Santa Cruz-BioE/Notebook
From 2014.igem.org
Important Protocols
Glycerol Stock Preparation
- Inoculated TSB with MR-1 (either from plate or glycerol stock) and grow overnight.
- add glycerol to culture to final concentration of 80% glycerol.
- 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.