Team:Caltech/Protocols/Flow cytometry
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# Set up 10mL LB cultures with appropriate antibiotic and aTc concentrations. Grow overnight at 37C while shaking. | # Set up 10mL LB cultures with appropriate antibiotic and aTc concentrations. Grow overnight at 37C while shaking. | ||
# 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. | # 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. | ||
# Spin down cultures in 15mL falcons at 10min, 3000xg and 22C. | # Spin down cultures in 15mL falcons at 10min, 3000xg and 22C. | ||
- | # Resuspend in 2mL [[Caltech/Protocols/BioAssay_Buffer|BioAssay Buffer]] (BAB). | + | # Resuspend in 2mL [[Team:Caltech/Protocols/BioAssay_Buffer|BioAssay Buffer]] (BAB). |
# Spin 2 min to pellet. | # Spin 2 min to pellet. | ||
# Resuspend in BAB + antibiotic + aTc (at same antibiotic and aTc concentrations as the original overnight cultures). | # Resuspend in BAB + antibiotic + aTc (at same antibiotic and aTc concentrations as the original overnight cultures). | ||
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# Put in 37C shaker for 4 hours. | # Put in 37C shaker for 4 hours. | ||
# Measure fluorescence of 1.5mL samples. | # Measure fluorescence of 1.5mL samples. | ||
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Latest revision as of 20:38, 8 September 2008
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