IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/06/05

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Results of Transformation

  • DB3.1 cells transformed with Chlor Construction Plasmid had 25 colonies growing on them. However there was a lawn of cells growing around the outside of the plate, possibly due to the plates not being spread properly with chloramphenical. Will streak two colonies on another properly spread chloramphenical plate to ensure that the cells have taken up the plasmid and become resistant
  • The BW27783 cells that were transformed with pUC19 had two colonies on a Amp plate, however both colonies had a couple of satellite colonies. As a result the Amp may be starting to degrade and the cells are not very competent. Will make new BW27783 competent cells.

Sequencing Results

  • 5 Biobricks (Amp construciton plasmid, GFP, J23100+RBS, pBAD, Terminator) that have been minipreped were sent for sequencing at NAPS. J23100+RBS was the only sequence that did not come back as a postive result. J23100+RBS was a weak signal, probably because it is a short sequence and hard to get a reading for

Bacterial Growth Curve

  1. We are doing this growth curve again because the previous one failed.
  2. Because we forgot to do an ON culture to innoculate from, we will instead innoculate directly from freezer stocks and measure OD as we go along.
  3. Today, we will also be using the Finlay Lab spec rather than the Lagally Lab sepc.
  4. Steps taken:
    1. Cleaned out cuvettes (and on the sides) with Kimwipes
    2. Set AMBL shake/incubator to 30ºC
    3. Poured 3x50 mL aliquots of LB broth into 3 flasks (50 mL each)
    4. Pipetted 1 mL LB into "Blank" cuvette
    5. Innoculated all 3 E. coli strains into flasks of LB (one strain per flask)
    6. Took OD readings for T=0 (9:30 AM)
  5. Steps for OD readings
    1. Pipetted 1 mL culture or LB into cuvette (LB for blank)
    2. Blanked spec with LB
    3. Take readings for all 3 samples
    4. Repeat steps 2 and 3
    5. Dispose of cells in biohazard liquid waste and rinse with ddH2O

Raw Data

Time Strain OD600 (1) OD600 (2) Average OD600 (2)
3:35 PM BW27783 0.126 0.130 0.128
4:07 PM BW27783 0.211 0.213 0.212
4:25 PM BW27783 0.247 0.253 0.250
4:45 PM BW27783 0.340 0.344 0.342
5:10 PM BW27783 0.476 0.478 0.477
5:40 PM BW27783 0.587 0.592 0.590
5:50 PM BW27783 0.625 0.622 0.624
6:00 PM BW27783 0.685 0.682 0.684
6:10 PM BW27783 0.741 0.737 0.739
6:40 PM DB3.1 0.114 0.112 0.113
7:00 PM DB3.1 0.142 0.145 0.144
7:20 PM DB3.1 0.151 0.154 0.153
7:40 PM DB3.1 0.228 0.228 0.228
8:10 PM DB3.1 0.296 0.295 0.296
8:40 PM DB3.1 0.396 0.397 0.397
9:15 PM DB3.1 0.467 0.466 0.467
9:45 PM DB3.1 0.555 0.558 0.557
9:55 PM DB3.1 0.578 0.584 0.581
10:05 PM DB3.1 0.603 0.601 0.602
10:15 PM DB3.1 0.626 0.625 0.626
10:30 PM DB3.1 0.663 0.660 0.662
7:40 PM DH5a 0.122 0.122 0.122
8:10 PM DH5a 0.157 0.155 0.156
8:40 PM DH5a 0.206 0.206 0.206
9:15 PM DH5a 0.251 0.255 0.253
9:45 PM DH5a 0.319 0.322 0.321
10:15 PM DH5a 0.392 0.394 0.393
10:30 PM DH5a 0.437 0.435 0.436
10:55 PM DH5a 0.488 0.484 0.486
11:30 PM DH5a 0.591 0.591 0.591
11:40 PM DH5a 0.668 0.666 0.667