User:Anand Gupta/Notebook/Microbial Fuel Cell/2009/04/28

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Overnight Culture Protocol

Overnight cultures are prepared in order to accumulate large numbers of bacteria in a liquid medium, which is useful in maximizing bacterial product yield. In the MFC, we'll be using liquid media at the Anode to allow for H+ ion flow as well as maximum respiratory yield(which will supply electrons for the cathode reduction).

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Protocol 2 Materials

  • LB Broth Powder
  • dH20
  • 500ml Flasks
  • E Coli k-12 stock plate
  • 15ml Sterile Tubes
  • Inoculating loops
  • Incubating Shaker
  • Alcohol/Chlorox [disinfectant]
  • Bunsen Burner/Bacti-incinerator

LB Broth Prep


1) Add 6.66g of LB Broth powder to 300ml of dH2O in autoclavable flask, stirring and heating to dissolve.

2) Autoclave flask for ~3hrs

Culture Prep


1) Add 10ml LB Broth to sterile tube.

2) Touch 1 colony on stock plate with sterile inoculating loop.

3) Place inoculating loop in Sample broth and swirl to release bacteria from loop

4) Incubate bacteria at 37C in shaker for ~24hrs.