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

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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

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