IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/05/26: Difference between revisions

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* Heat shock at 42.5ºC for 45 seconds
* Heat shock at 42.5ºC for 45 seconds
* 2 minutes on ice
* 2 minutes on ice
* Added 400 µL of sterile LB broth to the 100 µL of glycerol stocks (in the same tube as the glycerol stocks)
* Added 400 µL of sterile LB broth to the 100 µL of glycerol stocks (in the same tube as the glycerol stocks). Total volume in microfuge tube should be 500 µL.
* Recovery for 2 hours at 37ºC
* Recovery for 2 hours at 37ºC
* Spread plated 100 µL of Tet-Construction Plasmid (Tet-Con) cells onto Tetracycline plates, and did the same for Kan-Con plates.


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Revision as of 14:24, 26 May 2009

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Testing Chloramphenicol Plates

  • Cells grew in both LB-only and LB+Chlor
  • Possibly due to insolubility of antibiotic.
  • Made LB Broth only (10 mL) and LB Broth + Chloramphenicol (10 mL Broth + 10 µL Chloramphenicol)
  • Innoculated with DB3.1 cells
  • Left to grow in Lagally Lab shaker till next morning.

Obtained results on Tetracycline plate

  • retrieved plates at 13:06 from the incubator from Lagally Lab BioHazard room
  • observed no growth on LE-tet plate
  • observed cell growth on LE-only plate
  • Conclusion: Tetracycline is effective

Transformation of Biobricks

  • The GFP, pBAD, Terminator and J23100/RBS showed slow growth colonies, only becoming apparent this morning.
  • The Tet Construction Plasmid and Kan Construction Plasmid showed no growth, will perform another transformation of these Biobricks

Transformation of Tet and Kan Construction Plasmids

  • Transformation performed in DB3.1 cells
  • Tetracycline and Kanamycin construction plasmids - 1 µL of DNA into competent cells stock
  • On ice for 30 minutes
  • Heat shock at 42.5ºC for 45 seconds
  • 2 minutes on ice
  • Added 400 µL of sterile LB broth to the 100 µL of glycerol stocks (in the same tube as the glycerol stocks). Total volume in microfuge tube should be 500 µL.
  • Recovery for 2 hours at 37ºC
  • Spread plated 100 µL of Tet-Construction Plasmid (Tet-Con) cells onto Tetracycline plates, and did the same for Kan-Con plates.


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