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