IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/06/04

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Competence Test for BW27783 cells

  • Take out 100uL glycerol stocks of BW27783 and thaw on ice
  • Add 2uL of pUC19 to the BW27783 cells and keep on ice for 30min
  • Heat shock at 41.7°C for 60sec
  • Put on ice for 2min
  • Recover cells in 37°C incubator for 2hours
  • Spread plate cells on Amp plate

Transformation of Chloramphenicol Construction Plasmid

  • Take out 100uL glycerol stocks of DB3.1 from -80 freezer and thaw on ice
  • Add 1uL of Chlor Construction Plasmid to DB3.1 cells and keep on ice for 30min
  • Heat shock at 41.7 for 60sec
  • Put on ice for 2 min
  • Recover cells in 37°C incubator for 2 hours
  • Spread plate 500uL on Chlor plate

Results of Chloramphenicol Testing

  • Plates were taken from the Lagally Lab 37ºC incubator at 10:05 am.
  • Results:
Plate Condition DB3.1 Growth? DH5a Growth?
Chlor 0LB-onlyYesYes
Chlor 1LB + 25µg/mL ChloramphenicolNoNo

Comments

  • Obviously, the chloramphenicol works. Now, I will aliquot Paul's batch into 1 mL aliquots
  • Discarded old batch of Chlor-25 into biohazardous waste.
  • Leftover stock kept in -20ºC freezer
  • To help Heather with transformation of Chloramphenicol construction plasmid, I will spread plate 5 chlor plates with 25 µL of Chlor-25 stock on each plate.



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