IGEM:MIT/2006/Notebook/2007-7-25

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Things to Do

1. Check on gel- DID NOT WORK, WILL HAVE TO REDO WHEN MORE GELS COME IN

2. Run time course for smelling J45120 and J45181- WORKING ON IT

3. GC extract J45900 + IPTG- DONE

4. Check on cultures grown in Tom's media- DID NOT GROW (NEED TO CONSULT WITH TOM)

5. Smell cultures with known amounts of methyl salicylate (0 ppm, 1 ppm, 2.5 ppm, 5 ppm, 7.5 ppm, 10 ppm)- BARRY AND I THINK THAT THE SMELL THRESHOLD IS SOMEWHERE BETWEEN 1 PPM and 2.5 PPM AFTER INTENSE SMELLING)

6. Contact Tom about remaking media- DONE (awaiting reply)

7. Consult with Drew about the abstract- WILL DO TOMORROW

8. Redilute cultures and present 0 ppm (green), 0.5 ppm (red), and 1 ppm (blue), 2.5 ppm (pink), and 5 ppm (orange) MS to people in group meeting- WILL DO TOMORROW

9. Bring J45900+ sample to Alex- DONE

10. Restreak master plates that need restreaking- DONE

11. Set up B0015, J45995, J45996, R0040.E0840 cultures for plate reader experiment tomorrow- DONE

12. Make fresh LCs of J45995, J45996, J45120, & J45181 for miniprepping tomorrow- DONE

Smellathon

J45120

Time (Hrs)----OD600-----Smell----Noses

5------------0.16-------No, No----Me, Jason

6------------0.42-------No, No, Faint----Me, Barry, Jason

7------------0.76-------No, No, No----Me, Barry, Jason

8------------1.46-------Faint, Faint, Faint----Me, Barry, Jason

9------------1.72-------Faint, Faint, Faint----Me, Barry, Jason

J45181

Time (Hrs)----OD600-----Smell----Noses

5------------0.20-------No, No----Me, Jason

6------------0.51-------Faint, Faint, Faint----Me, Barry, Jason

7------------1.10-------Faint, Faint, Faint----Me, Barry, Jason

8------------1.58-------Stronger, Stronger, Stronger----Me, Barry, Jason

9------------1.86-------Stronger, Stronger, Stronger----Me, Barry, Jason

Conclusion: GC results were correct. For whatever reason, J45181 is producing more methyl salicylate in exponential phase than J45120. To sort out this situation, J45120, J45181, J45995, and J45996 will be miniprepped and sequenced before 10 AM tomorrow. Plate reader experiment is set for 9 AM tomorrow, so I must come in around 8 AM.

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