IGEM:MIT/2006/Notebook/2007-8-15

J45120/J45181 Time Course

Used protocol in yesterday's notebook entry

OD600s before salicylic acid addition

J45120

Hour 4- 0.22

Hour 5- 0.36

Hour 6- 0.90

Hour 7- 1.50

Hour 8- 1.96

Hour 9- 2.30

Hour 10- 2.48

Hour 11- 2.66

Hour 12- 2.68

J45181

Hour 4- 0.16

Hour 5- 0.36

Hour 6- 0.72

Hour 7- 1.20

Hour 8- 1.76

Hour 9- 2.28

Hour 10- 2.70

Hour 11- 2.60

Hour 12- 2.62

OD600s after salicylic acid addition

J45120

Hour 7- 1.02

Hour 8- 1.36

Hour 9- 1.70

Hour 10- 2.00

Hour 11- 2.18

Hour 12- 2.36

Hour 13- 2.56

Hour 14- 2.64

Hour 15- 2.74

J45181

Hour 7- 0.80

Hour 8- 1.24

Hour 9- 1.62

Hour 10- 1.96

Hour 11- 2.26

Hour 12- 2.30

Hour 13- 2.48

Hour 14- 2.48

Hour 15-

Other Things to Do

1. Ask Isadora to order more 125-mL flasks- DONE

2. Ask Drew if it's okay to be late for lab meeting- DONE (OKAY)

3. Bring GC column to Stephanopoulos lab- DONE

4. Make GC reservations for tomorrow- DONE

5. Set up fluorescent experiment following the procedure below- DONE

• Grow up 5-mL B0015, R0040.E0840, J45995, and J45996 cultures for 17 hours- DONE

• Dilute each culture 1:250 in a 25-mL culture in a 250-mL baffled flask- DONE

• Grow these cultures for 4 hours at 220 RPM at 37C- DONE

• Take OD600s of each culture: J45995- 1.28, J45996- 1.22, B0015- 0.88, R0040.E0840- 0.64- DONE

• Dilute the cultures: 2.5 mL for J45995 (22.5 mL media), 2.62 mL for J45996 (22.4 mL media), 3.64 mL for B0015 (21.4 mL media), 5 mL for R0040.E0840 (20 mL media) (all in 250-mL baffled flasks)- DONE

• Grow the cultures at 220 RPM for 50 mins- DONE

• Remove 200 uL triplet samples from each culture to run on the plate reader- DONE