"
Team:Chiba/Notebook/Calendar/23 September 2009
From 2009.igem.org
(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.
 2009 iGEM Chiba |
 face |
 Mr. M |
 pugyaaa |
 aiueo |
 using throwaway chopsticks |
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) |
Pictures are here.
photo
10:30 Start
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
11:00
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
11:30
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
12:00
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
12:30
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
13:00
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
13:30
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
14:00
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
14:30
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
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.
