IGEM:MIT/2006/Notebook/2007-8-24: Difference between revisions
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Hour 4- 0.06 | Hour 4- 0.06 | ||
Hour 5- | Hour 5- 0.16 | ||
Hour 6- | Hour 6- 0.48 | ||
Hour 7- | Hour 7- 1.00 | ||
Hour 8- | Hour 8- 1.56 | ||
Hour 9- | Hour 9- 1.94 | ||
Hour 10- | Hour 10- 2.18 | ||
Hour 11- | Hour 11- 2.38 | ||
Hour 12- | Hour 12- 2.42 | ||
J45181 | J45181 | ||
Hour 4- | Hour 4- 0.08 | ||
Hour 5- | Hour 5- 0.20 | ||
Hour 6- | Hour 6- 0.54 | ||
Hour 7- | Hour 7- 1.10 | ||
Hour 8- | Hour 8- 1.76 | ||
Hour 9- | Hour 9- 2.10 | ||
Hour 10- | Hour 10- 2.28 | ||
Hour 11- | Hour 11- 2.40 | ||
Hour 12- | Hour 12- 2.44 | ||
'''After Salicylic Acid Addition''' | '''After Salicylic Acid Addition''' | ||
| Line 71: | Line 71: | ||
J45120 | J45120 | ||
Hour 7- | Hour 7- 0.70 | ||
Hour 8- | Hour 8- 1.12 | ||
Hour 9- | Hour 9- 1.38 | ||
Hour 10- | Hour 10- 1.64 | ||
Hour 11- | Hour 11- 1.92 | ||
Hour 12- | Hour 12- 2.26 | ||
Hour 13- | Hour 13- 2.24 | ||
Hour 14- | Hour 14- 2.32 | ||
Hour 15- | Hour 15- 2.46 | ||
J45181 | J45181 | ||
Hour 7- | Hour 7- 0.72 | ||
Hour 8- | Hour 8- 1.16 | ||
Hour 9- | Hour 9- 1.56 | ||
Hour 10- | Hour 10- 1.80 | ||
Hour 11- | Hour 11- 1.94 | ||
Hour 12- | Hour 12- 2.14 | ||
Hour 13- | Hour 13- 2.28 | ||
Hour 14- | Hour 14- 2.36 | ||
Hour 15- | Hour 15- 2.54 | ||
=Fluorescence= | |||
Also set up 3 LCs of each J45995, J45996, R0040.E0840, and B0015 at 8:50 PM and made reservations for the plate reader (5 PM Saturday to 5 PM Sunday) | |||
=GC Results for Cumulative Methyl Salicylate= | =GC Results for Cumulative Methyl Salicylate= | ||
Latest revision as of 21:20, 24 August 2007
Redo of J45120/J45181 3-Hour Pulse
1. Add the following in a 2-L flask:
- 275 mL LB AMP/TET
- 27.5 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:
- 275 mL LB AMP/TET
- 27.5 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
OD600s
Before Salicylic Acid Addition
J45120
Hour 4- 0.06
Hour 5- 0.16
Hour 6- 0.48
Hour 7- 1.00
Hour 8- 1.56
Hour 9- 1.94
Hour 10- 2.18
Hour 11- 2.38
Hour 12- 2.42
J45181
Hour 4- 0.08
Hour 5- 0.20
Hour 6- 0.54
Hour 7- 1.10
Hour 8- 1.76
Hour 9- 2.10
Hour 10- 2.28
Hour 11- 2.40
Hour 12- 2.44
After Salicylic Acid Addition
J45120
Hour 7- 0.70
Hour 8- 1.12
Hour 9- 1.38
Hour 10- 1.64
Hour 11- 1.92
Hour 12- 2.26
Hour 13- 2.24
Hour 14- 2.32
Hour 15- 2.46
J45181
Hour 7- 0.72
Hour 8- 1.16
Hour 9- 1.56
Hour 10- 1.80
Hour 11- 1.94
Hour 12- 2.14
Hour 13- 2.28
Hour 14- 2.36
Hour 15- 2.54
Fluorescence
Also set up 3 LCs of each J45995, J45996, R0040.E0840, and B0015 at 8:50 PM and made reservations for the plate reader (5 PM Saturday to 5 PM Sunday)
GC Results for Cumulative Methyl Salicylate
J45120
Hour 15- 175.9 ppm
Hour 16- 151.5 ppm
Hour 17- 129.6 ppm
J45181
Hour 15- 226.3 ppm
Hour 16- 123.5 ppm
Hour 17- 189.3 ppm
Weird, huh?
For the time being, will try repeating 3-hour pulse experiment
