IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week3/Widgetry

Goals for this week

1. Make Orianna's growth media
2. construct more chambers to run parallel tests
3. set up Keithley for multiple tests
4. grow mtrA, mtrB,wt, E. coli in liquid cultures
5. negative control E. coli in LB/lactate
6. standardize tests (OD, bubbling, etc)

Tuesday: June 8, 2008

• Constructed two new MFCs
• grew mtrA, mtrB, wt, and E. coli in LB+lactate liquid cultures
• Ran negative control with E. coli in LML media

Wednesday: June 9, 2008

• Made Orianna's growth media

First followed recipe exactly: pH = 3.4

• Added NaOH to neutralize media

Missing amino acids

• Grew wt Shewie, mtrA, mtrB, and mtrC/omcA in Orianna's growth media
  • overnight cultures
  • RESULTS:

OD readings:

wt: -0.001

mtrA: 0.001

mtrB: 0.003

mtrC/omcA: 0.002

CONCLUSION:

Orianna's growth media isn't working. Go see Colleen for possible sources of error.

• OD readings from liquid cultures grown in LB + lactate growth media
  • wt: 2.489
  • mtrB: 1.711

• Ran MFC overnight with mtrB and wt Shewie strains.
  • added strains first without lactate
  • waited until chambers became anaerobic (color-change) to add lactate
  • wt cell chamber denser than mtrB cell chamber
  • N2 stopped in wt chamber. turned off N2 in mtrB chamber to mirror this change

• Experiment continued
  • replenished lactate in wt chamber
  • fluctuations may have resulted from air/N2 flow

Thursday: June 10, 2008

• Continued running experiment from Wednesday
• Purchased pipe cleaners, resistors
• Made OGM with potassium PIPES

Friday: June 11, 2008

Lab

• set up three chamber system
• tested wt, mtrA, negative control:
  • resistances:
    • wt:
    • mtrA:
    • none:

465 s: injected Shewie 3500 s: injected Lactate & stopped N₂