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