IGEM:MIT/2006/Notebook/2007-7-3: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(New page: =Things to Do= 1. Request Japanese plasmid which overexpresses leucine in E. coli 2. Ask Professor Prather if she thinks that leucine is limiting in our reaction 3. Perform experiment t...)
 
 
(25 intermediate revisions by the same user not shown)
Line 1: Line 1:
=Things to Do=
=Things to Do=


1. Request Japanese plasmid which overexpresses leucine in E. coli
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
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
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 Knight/Endy/grad students if they think that it is a good idea to add new promoters/RBSs to precursor devices in parallel with obtaining overexpressing strains
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)
 
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.

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.

Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:MIT/2006/Notebook/2007-7-3&oldid=127517"