IGEM:MIT/2006/Notebook/2007-1-31

Plan

1. Make reservations for the plate reader- DONE

2. Make reservations for the GC (isoamyl acetate samples)- DONE

3. Dilute the osmY and control cultures and grow them up for 3 hours before running on the plate reader- DONE

4. Run the osmY and control cultures on the plate reader- DONE

5. Submit the plates streaked out last night to the Registry- NOT DONE (missed Meagan twice today; will do it tomorrow)

6. Finish extracting the isoamyl acetate cultures- DONE

7. Run the isoamyl acetate cultures on the GC- DONE

8. Update google doc for the meeting- DONE

9. Get organized for the meeting at 4 PM- DONE

10. Move initial plate reader results to the wiki for the meeting- DONE

11. Go to the meeting at 4 PM on the status of the research- DONE

12. Analyze GC results- THINK MACHINE MAY HAVE BEEN SCREWY (CHANGED A PART AND RERUNNING IT)

13. Digest J45180 individually with SpeI, XbaI, and EcoRI overnight- DONE

14. Make LCs of J45600 hopefuls to smell tomorrow- DONE

Burning Questions

1. Do we still want to have the control mechanisms in the final system?

• If the samples on the GC test positively for isoamyl acetate, we have successfully created autonomous mint and banana cells.

• If there was not really a tube mix up and J45998 does in fact act exactly like R0040.E0840 due to an imbalance in the strength of the short osmY promoter and R0040, we will have to a) verify that the inverter.BSMT generator is present in the Registry and correct, b) ligate it to the long osmY promoter, and c) ligate the resultant piece to J45400 to make J45600.

2. How will we go about continuing the project?

• This semester probably will not be a good time for me to work on the project. I do not know if any other members of the team from last summer can work on it during the semester. I will be happy to work on it this summer if need be and still strongly believe that we should get this work published whether it be the constitutive or controlled version of the system.

3. What is left to do?

Answer for constitutive system: GC detection and analysis to find the exact concentrations of methyl salicylate and isoamyl acetate being produced. Then, there is the paper...

Answer for controlled system: Assuming that the short osmY promoter/inverter complex works, J45400 and J45250 must be ligated together to make J45600, and J45320 and J45180 must be ligated together to make J45800. Once these plasmids are cotransformed, a time course of samples from the cell's culture must be run on the GC. Then, there is the paper...