Team:Caltech/Protocols/Flow cytometry

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Latest revision as of 20:38, 8 September 2008


iGEM 2008



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Flow Cytometry


  1. Set up 10mL LB cultures with appropriate antibiotic and aTc concentrations. Grow overnight at 37C while shaking.
  2. Take out cultures and measure OD at 600nm. If it's below 0.4, place back at 37C for another 3 hours until it reaches OD ~0.5.
  3. Spin down cultures in 15mL falcons at 10min, 3000xg and 22C.
  4. Resuspend in 2mL BioAssay Buffer (BAB).
  5. Spin 2 min to pellet.
  6. Resuspend in BAB + antibiotic + aTc (at same antibiotic and aTc concentrations as the original overnight cultures).
  7. Measure OD and make sure it's between 0.4 and 0.6. If it's below 0.4, spin and pellet and try again.
  8. Put in 37C shaker for 4 hours.
  9. Measure fluorescence of 1.5mL samples.
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