IGEM:MIT/2006/Notebook/2007-8-23: Difference between revisions
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3. Grow both cultures at 110 RPM at 37C, starting J45200 20 minutes before J45250. Once the culture is put at 110 RPM and 37C, it is Time Zero | 3. Grow both cultures at 110 RPM at 37C, starting J45200 20 minutes before J45250. Once the culture is put at 110 RPM and 37C, it is Time Zero | ||
4. After x hours (for x=4 to x=12), extract 25 mL culture from master culture, pour into 125-mL flask, and add 0.5 mL .1 M salicylic acid to it for both J45120 and J45181 (grow up at 110 RPM) | |||
5. At hour x+3 for hour x, GC extract culture | |||
Revision as of 03:09, 23 August 2007
J45200/J45250 3-Hour Pulse experiment
Protocol
Protocol for 3-hour pulse:
1. Add the following in a 2-L flask:
- 275 mL LB AMP/TET
- 27.5 uL grown J45120 culture
2. Add the following in a 2-L baffled flask:
- 275 mL LB AMP/TET
- 27.5 uL grown J45181 culture
3. Grow both cultures at 110 RPM at 37C, starting J45200 20 minutes before J45250. Once the culture is put at 110 RPM and 37C, it is Time Zero
4. After x hours (for x=4 to x=12), extract 25 mL culture from master culture, pour into 125-mL flask, and add 0.5 mL .1 M salicylic acid to it for both J45120 and J45181 (grow up at 110 RPM)
5. At hour x+3 for hour x, GC extract culture
