IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/06/02: Difference between revisions

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*Put into Lagally Lab shaker @90rpm and 37°C
*Put into Lagally Lab shaker @90rpm and 37°C


==Chloramphenicol Testing
===Today's Test===
* Paul's new batch of chloramphenicol from his lab.
* Freshly made chloramphenicol stocks.
===Steps performed===
* Made chloramphenicol stocks (1000X) at 25 mg/mL
** 250.2 mg of chloramphenicol
** 10 mL of MeOH (99.8% purity)
** Filter sterilized in 0.45 µm polyethersulfone sterile syringe filter from VWR.
**


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Revision as of 15:07, 15 June 2009

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Results of Chloramphenicol-100 Test

  • Chloramphenicol plates at 100 µg/mL still did not work.
  • Growth observed on both LB-only and LB-chlor(100) plates, for both DB3.1 and DH5alpha cells.
  • Notes:
    • These plates were made using the old batch of chloramphenicol that Calvin had made, not his newer batch.
    • Will try spread plating Paul's batch of chloramphenicol.

Making Competent Cells

Checked the OD of the cells at 1220 (aim is to get A = [0.04,0.06])

  • BW27781 A = 0.024
  • DH5α A = 0.014
  • Conclusion: Desired concentration not yet reached
  • Put cells back into incubator

Checked the OD of the cells again at 1340

  • BW27781 A = 0.084
  • DH5α A = 0.024
  • Conclusion: Concentration of BW27781 passed the desired amount. DH5α still not ready
  • Put BW27781 into AMBL fridge to slow growth
  • Put DH5α back into incubator

Checked OD of DH5α at 1430

  • A = 0.112
  • Conclusion: Concentration passed the desired amount.
  • Put DH5α into AMBL fridge


  • Since the concentrations of both strains have passed the accepted value, the experiment has been terminated.
  • It was observed that many cuvettes are either dirty or scratched up. This will affect the accuracy of the OD reading. All cuvettes that are dirty or scratched up will be marked to prevent future inacurate reading.
  • To increase the efficiency of making future competent cells, a growth curve will be created for each strain (BW27781, DH5α and DB3.1).


Creating Growth Curve

  • Inoculated the 3 strains of cells (DB3.1, DH5α, BW27781) at 1520 in the Lagally Lab
  • Used 50mL sterile LB + cells (frozen glycerol stock -80°C)
  • Put into Lagally Lab shaker @90rpm and 37°C

==Chloramphenicol Testing

Today's Test

  • Paul's new batch of chloramphenicol from his lab.
  • Freshly made chloramphenicol stocks.

Steps performed

  • Made chloramphenicol stocks (1000X) at 25 mg/mL
    • 250.2 mg of chloramphenicol
    • 10 mL of MeOH (99.8% purity)
    • Filter sterilized in 0.45 µm polyethersulfone sterile syringe filter from VWR.


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