Team:Harvard/Dailybook/Week8/Widgetry

=Goals for this week=


 * 1) Run co-culture test with DH5α, DH5α/pUC19, and Shewie
 * 2) Move chambers to the "dark" to prevent a reaction from happening with light and oxygen
 * 3) Test light-repressible system on plates

=Monday: August 11, 2008=


 * cleaned chambers
 * moved setup to a cabinet to have system in the "dark"

DH5α, DH5α/pUC19, and Shewie co-culture experiment setup

 * Chamber 1: DH5α + wt Shewie + Lactose
 * Chamber 2: DH5α + wt Shewie + Lactate           (pos. control)
 * Chamber 3: DH5α + wt Shewie                     (neg. control)
 * Chamber 4: DH5α/pUC19 + wt Shewie + Lactose
 * Chamber 5: DH5α/pUC19 + wt Shewie + Lactate     (pos. control)
 * Chamber 6: DH5α/pUC19 + wt Shewie               (neg. control)
 * Chamber 7: wt Shewie + Lactose                  (neg. control)
 * Chamber 8: DH5α + Lactose                       (neg. control)
 * Chamber 9: DH5α/pUC19 + Lactose                 (neg. control)

=Tuesday: August 12, 2008=

First Half



 * Notes:
 * At t = 5000 s, all cells added
 * At t = 90000 s, all injections done
 * At t = 155000 s, Lactose injections on Chambers 1 and 4

Second Half

 * Notes:
 * Computer restarted in middle of night, t=0 in second graph is 7 hours after end of first graph
 * At t = 925000 s,
 * Lactose injections on Chambers 2 and 5
 * Lactate injections on Chambers 3 and 6

Concerns

 * Repeatable current production in DH5(alpha) alone
 * Possible contamination?
 * Distinguishing features to tell current production of DH5(alpha) and DH5(alpha)/pUC19 apart
 * After first lactose injection, the current production of the two have noticeable difference in slope
 * However, initially after second lactose injection, the graphs overlap

=Wednesday: August 13, 2008=

RU1012 Light Repressible system experiment

 * Streaked plates.
 * All Cm/Amp plates with Xgal


 * Dark
 * Plate 1: RU1012 w/ plasmids
 * Plate 2: RU1012
 * Plate 3: Negative control (No bugs)
 * Light
 * Plate 1: RU1012 w/ plasmids
 * Plate 2: RU1012
 * Plate 3: Negative control (No bugs)

Results

 * RU1012 with Plasmid
 * [[Image: RU1012_plasmid.JPG | 600px]]
 * Notes:
 * Plate on left was grown under light
 * Plate on right was grown in the dark
 * Possible repression seen in bottom left corner of the plate, but seems leaky


 * RU1012
 * [[Image: RU1012_light.JPG | 300px]] [[Image: RU1012_dark.JPG | 300px]]
 * Notes:
 * Plate on left was grown under light
 * Plate on right was grown in the dark
 * Exhibited same behavior as RU1012 with plasmid


 * Negative Control
 * Nothing on plates for both light and dark


 * Conclusions
 * Hard to distinguish if it worked with streaked colonies
 * May work better with lawns

Co-Culture Experiment Follow-up

 * When disassembling the fuel cells, took the chamber media and spread them on Xgal plates
 * Way of determining if contamination occured
 * Controls are not expected to turn blue

=Thursday: August 14, 2008=

RU1012 Light Repressible system experiment

 * Spread plates to try to create lawn.
 * All Cm/Amp plates with Xgal


 * Dark
 * Plate 1: RU1012 w/ plasmids
 * Plate 2: RU1012
 * Plate 3: Negative control (No bugs)
 * Light
 * Plate 1: RU1012 w/ plasmids
 * Plate 2: RU1012
 * Plate 3: Negative control (No bugs)

Results

 * RU1012 with plasmid under light
 * [[Image: RU1012_plasmid_light.JPG | 300 px]]
 * Note:
 * Colonies had specks of blue, but not very noticeable


 * RU1012 with plasmid in the dark
 * [[Image: RU1012_plasmid_dark.JPG | 300 px]]


 * Conclusion
 * Possibly light is too strong? Explaining no growth in middle of plate grow under light

=Friday: August 15, 2008=

RU1012 Light Repressible system experiment

 * Repeated previous day's experiment exactly
 * Placed light further away

Co-culture Experiment Follow-up

 * All plates turned blue
 * Control chambers very likely were contaminated
 * Will explore possibilities of autoclaving chambers