IGEM:MIT/2006/Notebook/2007-8-14: Difference between revisions
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Protocol for 3-hour pulse: | Protocol for 3-hour pulse: | ||
1. Add the following in a 2-L flask: | 1. Add the following in a 2-L flask: | ||
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7. At hour x+3 for hour x, GC extract culture | 7. At hour x+3 for hour x, GC extract culture | ||
=Today's experiment= | |||
Time Zero= 10:53 (time at which master flask starts shaking at 110 RPM, wanted to do it earlier but the flask broke on the shaker and spent a good hour or so fixing it and then needed to redo cultures) | Time Zero= 10:53 (time at which master flask starts shaking at 110 RPM, wanted to do it earlier but the flask broke on the shaker and spent a good hour or so fixing it and then needed to redo cultures) |
Revision as of 07:56, 14 August 2007
Smell Course Set Up
Protocol for 3-hour pulse:
1. Add the following in a 2-L flask:
- 250 mL LB AMP/TET
- 25 uL grown J45120 culture
2. Grow the culture up for 20 mins at 220 RPM at 37C
3. Add the following in a 2-L baffled flask:
- 250 mL LB AMP/TET
- 25 uL grown J45181 culture
4. Grow the culture up for 20 mins at 220 RPM at 37C
5. Lower the RPM for both cultures to 110 RPM at 37C. The start of this step is Time Zero.
6. 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)
7. At hour x+3 for hour x, GC extract culture
Today's experiment
Time Zero= 10:53 (time at which master flask starts shaking at 110 RPM, wanted to do it earlier but the flask broke on the shaker and spent a good hour or so fixing it and then needed to redo cultures)
Hour 5= 3:53 (add salicylic acid to a culture)
Hour 6= 4:53 (add salicylic acid to a culture)
Hour 8= 6:53 (smell Hour 5 culture)
Hour 9= 7:53 (smell Hour 6 culture)
Hour 10.5= 9:23 (add salicylic acid to a culture)
Hour 11.5= 9:53 (add salicylic acid to a culture)
Hour 13.5= 12:23 (smell Hour 10.5 culture)
Hour 14.5= 1:23 (smell Hour 11 culture)