IGEM:MIT/2006/Notebook/2007-8-20: Difference between revisions
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2. Call for a meeting tomorrow- DONE | 2. Call for a meeting tomorrow- DONE | ||
3. Discuss with Veena and Drew | 3. Discuss conference logistics with Veena and Drew | ||
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). | 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). | ||
Revision as of 11:25, 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 conference logistics with Veena and Drew
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- DONE
7. Make Tris buffer solution- DONE
8. Ask Isadora about ordering more heptane
Protocol for Time Course
1. At each time point, aliquot 20 mL culture into falcon tube and remove .5 mL to take an OD600 measurement.
2. Put Tris buffer (10 mM Tris, 100 mM NaCl, pH 8) at 4C.
3. Incubate culture for 10 minutes on ice.
4. Centrifuge the sample at 4C for 4 mins at 13000 RPM (balancing it with a falcon tube containing 20-mL water).
5. Remove supernatant.
6. Resuspend sample pellet in 10 mL Tris buffer.
7. Repeat steps 4 and 5.
8. Resuspend cells in 1 mL B-PER II.
9. Add 19 mL water to sample.
10. Add .4 mL .1 M salicylic acid to sample.
11. Let sample incubate for 2 hours at room temperature after addition of salicylic acid.
12. GC extract the sample.
