Team:Chiba/Notebook/Calendar/23 September 2009
From 2009.igem.org
(→Examine limit of AHL generation) |
(→LuxR Mutantの個性決める実験の写真だけうp) |
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Image:Chiba_2309_1430_1000.jpg|(5)AHL 1000nM | Image:Chiba_2309_1430_1000.jpg|(5)AHL 1000nM | ||
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== Examine limit of AHL generation(1)-2 == | == Examine limit of AHL generation(1)-2 == |
Revision as of 04:56, 26 September 2009
(22_September_2009 <|>24_September_2009)
Contents |
E. coli Painting (2)
We draw various pictures with ink brush and throwaway chopsticks.
Yesterday's operation is here.
To judge character of LuxR mutants (2)-2
Yesterday's operation is here.
- Today's operation
10:30
We transplanted E.coli, which has been cultured on NC filter, to solid medium which contains each concentration of AHL.
AHL concentration is : 0, 1, 10, 100, and 1000 nM
We decided this time is T=0 and observed condition of fluorescence every 30 min.
- Mutants' Location
Mutant 1 x 3 well | Mutant 9 x 3 well |
Mutant 2 | Mutant 10 |
Mutant 3 | Mutant 11 |
Mutant 4 | Mutant L1 |
Mutant 5 | Mutant L2 |
Mutant 6 | Wild Type |
Mutant 7 | Negative Control |
Mutant 8 | (Nothing) |
Picture are here.
photo
10:30 Start
11:00
11:30
12:00
12:30
13:00
13:30
14:00
14:30
Examine limit of AHL generation(1)-2
Yesterday's operation is here.
- Today's operation
11:00
We did main culture.
22:00
We measured OD.
OD600=1.98
Then, we spun down this culture solution and add 12.5 mL of flesh LB-Amp.
22:10
We add 12.5 μL of IPTG to the culture solution and started culture at 37 degrees Celsius.
Transformation(2)-1
- Plasmid
plux-GFP(Cm, p15A)
- Competent Cells
JW1262(pCIA3-LuxR(Amp))
JW1262(Null)
21:35
We transplanted these bacteria each plates.
pCIA3-LuxR and plux-GFP --> LB-Amp, Cm plate
plux-GFP only --> LB-Cm plate
21:50-
We cultured these plates.