Talk:The BioBricks Foundation:Standards/Technical/Measurement/Promoter characterization experiment (FACS)

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LBL: Effects of Varying the Growth conditions on the Promoter characterization experiment

  • Because of my poor planning, we made the following modifications to the procedure, resulting in relatively minor differences in the ratio of GFP intensities:
    • Culturing the strains
      • grew in LB + Kan overnight
      • grew at 37C shaking at 225 rpm
      • back diluted 1:1000 (instead of 1:100 doh!!) supplemented M9 media, and grew for 4 hours so OD600<0.1
    • Live Cell FACS Procedure
      • washed only once with PBS
  • CAjoF 17:45, 5 March 2008 (EST)

MIT: Experimental details

  • We only spun-down and resuspended 1ml of each sample in PBS.
  • We also only did one wash with PBS, so I'm updating the protocol for future labs doing this -- since Caroline made the same choice :)
  • Our PBS is from invitrogen and looks like it has a different formulation then what caroline listed in materials.

VTech:Experimental details

  • Grew a single colony overnight and then split the culture into 3 tubes in the dilution in the morning, rather than starting 3 overnight cultures from 3 seperate colonies.
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