IGEM:MIT/2006/Notebook/2007-7-17

=To Do Today=

1. Taking OD600 readings from J45181 and J45120 cultures using new protocol to construct a growth curve- WORKING ON IT

2. Wait for the cells from last night to grow up a bit to exponential phase (check OD600s)- DONE

3. Run a protein gel of cells in exponential phase using the same protocol as last time- RUNNING (unfortunately, could not find molecular marker)

4. Discuss media with Dr. Knight- DONE

5. Continue to discuss with Li Li about the mechanics of the calibration curve- DONE

6. GC extract full-grown culture for extraction efficiency measurement tomorrow

7. Check sequencing of J45600 and J45800 again- do it tomorrow

8. Check secondary structures of precursor devices with Barry- DONE (One of them we may have to worry about)

9. Grow up J45800 (induced and not induced), J45700 (induced and not induced), and J45181 (control) on new media- DONE

10. AB3257 (aroF- aroG- aroH- strain) finally grew up. Make an LC of it tonight- DONE

=Growth Curve=

Hour--J45120--J45181

3---0.00--0.02

4---0.03--0.07

5---0.16--0.28

6---0.48--0.68

7---1.00--1.34

8---1.70--1.88

9---2.08--2.10

10--2.22--2.22

11--?--?

12--2.36--2.44

Conclusion- Will use same protocol proposed

=ODs for protein gel cultures=

600+ .89

600- .91

250 1.05

800+ 1.01

800- 1.16

181 1.05

B0015 1.01