IGEM:MIT/2006/Notebook/2007-6-13

Plan

1. J45400 did not grow up in any of the LCs!!! Uggh!! Contact Meagan about obtaining J45400 from the Registry's glycerol stock or an extra plate- DONE, will pick up a streaked plate this afternoon

2. Contact Drew about possibly doing a time course of J45250 and J45181 with the precursors added- DONE, said to ask Barry who said to go ahead with it

3. Contact list to see if anyone remembers how to make sodium salicylate solution (we have run out)- DONE (awaiting reply)

4. Heat shock overnight digests of J45181- DONE

5. Run 2 J45181 digests on gel and gel extract 1 J45181 digest- DID NOT CUT

6. Redo digest of J45181 with new enzymes (XbaI, PstI, + XbaI and PstI for each of the three)- DONE

7. Run on gel- DID NOT CUT AGAIN, looks like X isn't cutting although one of the single cuts looked cut --> will do overnight digests of J45181 ES & J45320 XP

7. Put freshly made plates away- DONE

8. Obtain Top 10 cells from Knight lab- DONE

9. Pick up streaked J45400 plate from Meagan & put in warm room- DONE *Theory on J45400- I think that the glycerol in our fridge was mislabeled as having the part in 1AK3 because the plate I received from Meagan was labeled as the part being on 3K3. This is possible because when I grew up the part last night, I grew it up on AK, AC, and AT but not just K.

10. Make LB Kan media- DONE

11. Make a 15 mL culture of the J45400 glycerol from our fridge in LB Kan media to test theory- DONE

12. Make a 5 mL culture of J45181 to do a time course of tomorrow early morning- DONE

13. Contact Li Li about using the GC/MS- will do tomorrow

14. Find a method for making more salicylic acid solution- DONE FINALLY

15. Add fresh salicylic acid solution + little left of original salicylic acid solution to 10 mL cultures of J45181 to make sure it smells before time course experiment

16. Make extra 15 mL culture of J45181 to digest with J45400 tomorrow (yes, it is growing up yippee)- DONE