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)

=Monday: June 7, 2008=

=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




 * Nitrogen stopped bubbling somewhere between 70,000 & 80,000