Penn State University 2006:Motility-assays: Difference between revisions
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===<font color="blue">Swarm Plate Assays</font>=== | ===<font color="blue">Swarm Plate Assays</font>=== | ||
Time: 2 hr first day; 9 hr second day | ''Time: 2 hr first day; 9 hr second day'' | ||
Reference: Garza, A.G., et al. J. Mol. Biol. (1996) 258, 270-285 | |||
''Reference: Garza, A.G., et al. J. Mol. Biol. (1996) 258, 270-285'' | |||
Media/Reagents | Media/Reagents | ||
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5. take picture of plate with digital camera | 5. take picture of plate with digital camera | ||
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[http://openwetware.org/wiki/IGEM:PennState Main] | |||
Revision as of 16:14, 24 October 2006
Swarm Plate Assays
Swarm Plate Assays
Time: 2 hr first day; 9 hr second day
Reference: Garza, A.G., et al. J. Mol. Biol. (1996) 258, 270-285
Media/Reagents LB Typtone broth plates Cells (from plate)
Day 1: Make Tryptone Swarm Plates 1. For each plate to be made, add the following to a 125mL flask: 0.25g Bacto tryptone 0.20g NaCl 0.15g sodium citrate 0.05g Eiken agar (for a 0.20% agar swarm plate) 2. add 25mL purified water to the flask and swirl to dissolve 3. autoclave 25 minutes with slow exhaust 4. upon cooling, add appropriate antibiotics and inducers - IPTG saturation concentration => 1mM - HSL saturation concentration => 100µM 5. allow plates to dry overnight
Day 1: Inoculate O’N culture
1. inoculate 3mL overnight cultures of all samples to be tested and shake at 200 RPM in 30°C incubator
Day 2: Assay 1. dilute overnight culture 1:100 in LB (30uL of culture in 3mL LB) 2. grow in a 30°C incubator at 200 RPM to an A590nm O.D. between 0.3-0.4 3. add about 3uL of each culture to plates 4. grow in a 30°C incubator 8 hours 5. take picture of plate with digital camera
