Revision as of 08:49, 21 August 2007

Things to Do

1. Early morning- run GC samples from last night- DONE

2. Prepare for meeting with Reshma

OD600s: MISSED 13 hours, J45120 14 hours- 2.48, J45181 14 hours- 2.46

1. At each time point, aliquot 20 mL cultures (J45120 and J45181) into falcon tubes and remove .5 mL to take an OD600 measurement.

2. Incubate culture for 10 minutes on ice.

3. Centrifuge the samples at 4C for 20 mins at 3800 RPM.

4. Discard the supernatants. Store the pellets at -80C until the last time point.

5. Once all time points are collected, resuspend pellets in 10 mL Tris buffer (10 mM Tris, 100 mM NaCl, pH 8).

6. Centrifuge the samples at 4C for 20 mins at 3800 RPM.

7. Discard supernatants.

8. Resuspend cells in 1 mL B-PER II.

9. Add 19 mL PBS to samples.

10. Add .4 mL .1 M salicylic acid to samples at staggering 20 min time points.

11. Let sample incubate for 2 hours at room temperature after addition of salicylic acid.

12. GC extract the samples.

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 OD600s: MISSED 13 hours, J45120 14 hours- 2.48, J45181 14 hours- 2.46
+. At each time point, aliquot 20 mL cultures (J45120 and J45181) into falcon tubes and remove .5 mL to take an OD600 measurement.
+. Incubate culture for 10 minutes on ice.
+. Centrifuge the samples at 4C for 20 mins at 3800 RPM.
+. Discard the supernatants.  Store the pellets at -80C until the last time point.
+. Once all time points are collected, resuspend pellets in 10 mL Tris buffer (10 mM Tris, 100 mM NaCl, pH 8).
+. Centrifuge the samples at 4C for 20 mins at 3800 RPM.
+. Discard supernatants.
+. Resuspend cells in 1 mL B-PER II.
+. Add 19 mL PBS to samples.
+. Add .4 mL .1 M salicylic acid to samples at staggering 20 min time points.
+. Let sample incubate for 2 hours at room temperature after addition of salicylic acid.
+. GC extract the samples.