Team:Groningen/Notebook/Wetwork 12September2012



Test GFP: reaction to rotten meat

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

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.


put promoters of LMU Munchen in dh5a

transformation of ~1 uL at 15:00
mistake! forgot to add LB medium!! --> Plate out anyway (amp plates) + O/N 5 mL culture of the rest..

O/N culture of:

  • rfp-psac-cm in dh5a: 2x from 9/09 (our competent cells)
  • rfp-psac-cm in dh5a: 5x from 10/09 (new competent cells)

at 15:00, 37 degrees C.


Another batch of competent cells in the making!


Another cloning for pSac-Cm+promoters+reporters and BBa_K823023+promoters+reporters in progress. We borrowed some tubes of E. coli DH5a competent cell from Sjoerd, our lovely supervisor.
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