IGEM:MIT/2006/Notebook/2007-8-16
Things today
1. Miniprep and sequence LC of J45250 1AT3- DONE (they said I'll most definitely get results by Monday morning, so I can hopefully start a banana time course; if not, I have the prepped DNA of the correct J45250 1AT3 whose LC I stupidly threw out, so I can transform that and start a new LC)
2. Data analysis of plate reader results- DONE (injector still broken; growth curves look like rainbows)
3. Run ALL 18 GC samples- DONE
4. Make the following cultures to prepare for GC tomorrow:
- 25-mL J45200 w/ precursor
- 25-mL J45250 (1AT3) w/ precursor
- 25-mL J45200 w/o precursor (control)
- 25-mL J45600
- 25-mL J45900
Calculation of Precursor
We initially forgot to take into account isoamyl alcohol's density when deciding at which concentration to add isoamyl alcohol.
We used 4.4 uL/10 mL because we assumed density to be 1 g/mL... Thus, .0044 g/.01 L*1 mol/88.2 g = 5 mM
To make the final concentration be 5 mM, we actually want 4.4 uL/.809 (g/mL density)= 5.44 uL.
Results from GC
J45120
4-7 Hours: 1.1 ppm
5-8 Hours: 13.7 ppm
6-9 Hours: 13.9 ppm
7-10 Hours: 40.7 ppm
8-11 Hours: 62.3 ppm
9-12 Hours: 67.6 ppm
10-13 Hours: 59.4 ppm
11-14 Hours: 66.6 ppm
12-15 Hours: 50.6 ppm
J45181
4-7 Hours: 7.9 ppm
5-8 Hours: 14.4 ppm
6-9 Hours: 27.3 ppm
7-10 Hours: 59.0 ppm
8-11 Hours: 80.6 ppm
9-12 Hours: 80.1 ppm
10-13 Hours: 63.4 ppm
11-14 Hours: 50.2 ppm
12-15 Hours: 37.7 ppm
