Team:Groningen/Notebook/Wetwork 12September2012

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Test GFP: reaction to rotten meat

meat: 5 gram aliquot from freezer, 3 days at 20 degrees Celsius

tested: Bacillus subtilis sp. 168 _ SboA_GFP in psac-cm (subjected to meat, and without meat) and WT (without meat)

11:00 inoculate O/N culture in SMM in normal 37 degrees stove.

  • OD SboA = 0.302
  • OD WT = 0.317

12:30 SboA-GFP to micro-array test setup, start OD = 0.1 (see notebook Wetwork 02/09 and 27/06). Also put SboA-GFP and WT in normal 37 degrees stove (start OD = 0.1).

14:30 harvest cells

  • OD WT = 1.168
  • OD SboA(no meat) = 1.023
  • OD SboA(meat) = 1.153


put cells on microscope slide for fluorescence microscopy:

  • agarose (1% in SMM) 500 uL on slide
  • spread with glass plate
  • spot 1 uL of sample on slide (duplo)

microscope results: no GFP, only autofluorescence.

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