Revision as of 10:57, 20 August 2007

Things to Do

1. Develop protocol for lysing cells (I see B-PER II solution on his bench)- DONE (have a protocol to follow, see below)

2. Call for a meeting tomorrow- DONE

3. Discuss with Veena and Drew about Conference

4. Do lysis protocol/wintergreen time course for Hour 13 (2:58, 3:18), 14 (3:58, 4:18), and 15 (4:58, 5:18).

5. Organize results from plate reader experiment and send it out to advisors- DONE

6. Collect materials for lysis step

7. Ask Isadora about ordering more heptane

1. At each time point, aliquot 20 mL culture into falcon tube and remove .5 mL to take an OD600 measurement.

2. Balance culture with 20 mL water in another tube in big centrifuge and put the settings at 4C, 10 mins, and 3000 RPM.

3. Remove supernatant and store the pellet at -80C until the last time point is taken.

4. Once all pellets are collected, follow steps 2-8 in lysis method 1.1 http://openwetware.org/wiki/Endy:Victor3_plate_reader/Calibrating_the_GFP-separated_label, adding 20X the amount of each reagant (10 mL Tris and 1 mL B-PER II) to the sample and performing all of the spinning steps in the Falcon tubes on the big centrifuge.

5. Add 19 mL LB AMP/TET to the samples.

6. Stagger addition of .4 mL .1 M salicylic acid by 20 minutes for each sample.

7. Let samples incubate for 2 hours after addition of salicylic acid.

8. GC extract the samples at the staggered time points.

@@ Line 25: / Line 25: @@
 . Once all pellets are collected, follow steps 2-8 in lysis method 1.1 http://openwetware.org/wiki/Endy:Victor3_plate_reader/Calibrating_the_GFP-separated_label, adding 20X the amount of each reagant (10 mL Tris and 1 mL B-PER II) to the sample and performing all of the spinning steps in the Falcon tubes on the big centrifuge.
-. Add 19 mL H20 to the samples.
+. Add 19 mL LB AMP/TET to the samples.
 . Stagger addition of .4 mL .1 M salicylic acid by 20 minutes for each sample.