IGEM:MIT/2006/Notebook/2007-7-3: Difference between revisions
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2. Ask Professor Prather if she thinks that leucine is limiting in our reaction- DONE | 2. Ask Professor Prather if she thinks that leucine is limiting in our reaction- DONE | ||
3. Perform experiment that Reshma suggested (add .02 g/L of leucine to J45600 and J45900 cultures) | 3. Perform experiment that Reshma suggested (add .02 g/L (25 uL of 20 g/L stock) of leucine to J45600 and J45900 cultures (25 mL))- DONE | ||
4. Resend request to Yale for aroF- aroG- aroH-, make sure it gets ordered today- DONE (AB3257 (aroF- aroG- aroH-) is on its way in today's mail!!!) | 4. Resend request to Yale for aroF- aroG- aroH-, make sure it gets ordered today- DONE (AB3257 (aroF- aroG- aroH-) is on its way in today's mail!!!) | ||
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5. Ask Endy if they think that it is a good idea to add new promoters/RBSs to precursor devices in parallel with obtaining overexpressing strains/what overall goals should be a priority- DONE | 5. Ask Endy if they think that it is a good idea to add new promoters/RBSs to precursor devices in parallel with obtaining overexpressing strains/what overall goals should be a priority- DONE | ||
6. Run GC samples from yesterday's time course- | 6. Run GC samples from yesterday's time course- DONE (see below) | ||
7. Meet with Alex about GC (2 PM)- DONE (need to prepare isoamyl acetate in heptane and heptane extraction of LB media, Alex seemed optimistic) | 7. Meet with Alex about GC (2 PM)- DONE (need to prepare isoamyl acetate in heptane and heptane extraction of LB media, Alex seemed optimistic) | ||
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3---------------------0.01------------NOT DETECTED------------------------------------------- | 3---------------------0.01------------NOT DETECTED------------------------------------------- | ||
5---------------------0.21------------(-0.4)------------------------------------------------- | 5---------------------0.21------------(-0.4)----------------------------------------------------------- | ||
7---------------------1.09------------2.6-----------------------2.39 | 7---------------------1.09------------2.6------------------------------------------2.39 | ||
9--------------------2.5--------------16.9----------------------6.76 | 9--------------------2.5--------------16.9-----------------------------------------6.76 | ||
11--------------------2.5------------- | 11--------------------2.5-------------12.8-----------------------------------------5.12 | ||
13--------------------2.8------------- | 13--------------------2.8-------------11.8-----------------------------------------4.21 | ||
'''J45181 (Exponential-Phase Sensitive Promoter)''' | '''J45181 (Exponential-Phase Sensitive Promoter)''' | ||
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3---------------------0.01------------NOT DETECTED------------------------------------------ | 3---------------------0.01------------NOT DETECTED------------------------------------------ | ||
5---------------------0.27------------(0.1)------------------------------------------------- | 5---------------------0.27------------(0.1)------------------------------------------------------------ | ||
7---------------------1.15------------6.5-----------------------5.65 | 7---------------------1.15------------6.5-----------------------------------------5.65 | ||
9--------------------2.0--------------14.0----------------------7.00 | 9--------------------2.0--------------14.0----------------------------------------7.00 | ||
11--------------------3.0------------- | 11--------------------3.0-------------11.5----------------------------------------3.83 | ||
13--------------------3.2------------- | 13--------------------3.2-------------4.0-----------------------------------------1.25 | ||
Thoughts for next time: | |||
Perhaps add in twice as many cells, so hour 5 is meaningful and hour 9 will likely be the beginning of stationary phase. | |||
Latest revision as of 15:08, 3 July 2007
Things to Do
1. Request Japanese plasmid which overexpresses leucine in E. coli- DONE
2. Ask Professor Prather if she thinks that leucine is limiting in our reaction- DONE
3. Perform experiment that Reshma suggested (add .02 g/L (25 uL of 20 g/L stock) of leucine to J45600 and J45900 cultures (25 mL))- DONE
4. Resend request to Yale for aroF- aroG- aroH-, make sure it gets ordered today- DONE (AB3257 (aroF- aroG- aroH-) is on its way in today's mail!!!)
5. Ask Endy if they think that it is a good idea to add new promoters/RBSs to precursor devices in parallel with obtaining overexpressing strains/what overall goals should be a priority- DONE
6. Run GC samples from yesterday's time course- DONE (see below)
7. Meet with Alex about GC (2 PM)- DONE (need to prepare isoamyl acetate in heptane and heptane extraction of LB media, Alex seemed optimistic)
8. Print out GC procedure for isoamyl acetate for meeting- DONE
Results from Time Courses
J45120 (Constitutive Promoter)
Time (Hrs)------------OD600-----------Methyl Salicylate (ppm)---Methyl Salicylate (ppm)/OD600
3---------------------0.01------------NOT DETECTED-------------------------------------------
5---------------------0.21------------(-0.4)-----------------------------------------------------------
7---------------------1.09------------2.6------------------------------------------2.39
9--------------------2.5--------------16.9-----------------------------------------6.76
11--------------------2.5-------------12.8-----------------------------------------5.12
13--------------------2.8-------------11.8-----------------------------------------4.21
J45181 (Exponential-Phase Sensitive Promoter)
Time (Hrs)------------OD600-----------Methyl Salicylate (ppm)---Methyl Salyclate (ppm)/OD600
3---------------------0.01------------NOT DETECTED------------------------------------------
5---------------------0.27------------(0.1)------------------------------------------------------------
7---------------------1.15------------6.5-----------------------------------------5.65
9--------------------2.0--------------14.0----------------------------------------7.00
11--------------------3.0-------------11.5----------------------------------------3.83
13--------------------3.2-------------4.0-----------------------------------------1.25
Thoughts for next time:
Perhaps add in twice as many cells, so hour 5 is meaningful and hour 9 will likely be the beginning of stationary phase.
