IGEM:MIT/2006/Notebook/2007-7-31

Set up for exponential phase extraction

• 25 uL overnight culture of J45181 in 250 mL LB A/T media (2-L flask)

• 25 uL overnight culture of J45120 in 250 mL LB A/T media (2-L flask)

• 2.5 uL overnight culture of J45800 in 25 mL LB A/K media (125-mL flask)

• 5 uL exponential-phase culture of B0015 in 25 mL LB A/K media (125-mL flask)

Grown up for 20 mins at 220 RPM, then switched to 110 RPM

J45120 6 hrs 0D600- 0.45, extract 6 mL, put at 4C for 2 hrs

J45181 6 hrs OD600- 0.67, extract 6 mL, put at 4C for 2 hrs

Other things to do

1. Phone meeting with Drew- DONE

2. Contact Alex about isoamyl acetate GC- DONE (awaiting reply)

3. Contact Dr. Tribe about media which will help overproduce chorismate in cells- DONE (awaiting reply)

4. Rework abstract for Amgen people- DONE

5. Rework Drew's edited abstract

6. Grow wild-type cells on M9 minimal media and see if they grow

7. Grow J45700, J45800, and wild-type cells on Neidhardt media

8. Do protein gel sample preparation for J45120 (6 hr growth, 7 hr growth, 8 hr growth), J45181 (6 hr growth, 7 hr growth, 8 hr growth), & B0015

9. At the end of the day, make LCs of J45995, J45996, B0015, R0040.E0840 to grow up for 16 hrs