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<li>① Set your instrument to read Abs600 (as OD calibration setting)</li> | <li>① Set your instrument to read Abs600 (as OD calibration setting)</li> | ||
<li>② Measure Abs600 of the overnight cultures</li> | <li>② Measure Abs600 of the overnight cultures</li> | ||
− | <li>③ | + | <li>③ Dilute the cultures to a target Abs600 of 0.02 (see the volume of preloading culture and media in Excel Dilution Calculation sheets) in 12 ml LB medium + Chloramphenicol in 50 mL falcon tube.</li> |
− | + | <li>④ Remove 500uL of each culture for your zero time point and hold these samples on ice. (You should have 16 sample tubes from the time=0 time point)</li> | |
− | <li> | + | <li>⑤ Incubate the cultures at 37°C and 220 rpm.</li> |
− | <li> | + | <li>⑥ Remove 500uL samples of each culture after 2, 4 and 6 hours of growth. Keep samples on ice while completing the additional time points. You should have 16 sample tubes for each time point.</li> |
− | <li> | + | <li>⑦ Set up your four measurement plates: For colony #1 culture, pipet 100 uL samples into wells A, B, C and D of the row for that device. For colony #2, pipet 100uL samples into wells E, F, G, and H of the row for that device.</li> |
− | <li> | + | <li>⑧ Read your plates, taking care to use the exact same settings used for your fluorescein measurement.</li> |
− | <li> | + | |
Revision as of 20:20, 31 October 2017