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Team:Chiba/Notebook/Calendar/25 September 2009
From 2009.igem.org
(24_September_2009 <|>26_September_2009)
Contents |
To judge character of LuxR mutants (3)-2
Yesterday's operation is here.
- Today's operation
10:45-
We transplanted E.coli by 48 pins to NC filter and cultured it.
22:20 measurement start
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.
22:20 Start
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
23:20
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
23:50
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
24:20
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
24:50
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
25:20
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
25:50
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
26:20
 (1) AHL 0 nM |
 (2) AHL 1 nM |
 (3) AHL 10 nM |
 (4)AHL 100nM |
 (5)AHL 1000nM |
Examine limit of AHL generation(2)-1
11:05-
We did prior culture.
glycerol stock : plux-GFP with pCIA3-LuxR
23:35-
We attenuated culture solution 1/105 and dispersed on plate.
Then we cultured this plate at 37 degrees Celsius.
And We made plates from each of mixture and 10 mL of LB-Amp, Cm solution medium.
- Element of mixtures
| Sample Namber | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
| supernatant solution | 5 mL | --- | --- | --- | --- | --- | --- | 200 μL |
| 1 | --- | 5 mL | --- | --- | --- | --- | --- | --- |
| 2 | --- | --- | 5 mL | --- | --- | --- | --- | --- |
| 3 | --- | --- | --- | 5 mL | --- | --- | --- | --- |
| 4 | --- | --- | --- | --- | 5 mL | --- | --- | --- |
| 5 | --- | --- | --- | --- | --- | 5 mL | --- | --- |
| 6 | --- | --- | --- | --- | --- | --- | 5 mL | --- |
| LB-Amp,Cm(liquid) | 10 mL | 10 mL | 10 mL | 10 mL | 10 mL | 10 mL | 10 mL | 9.8 mL |
| Total | 10 mL | 10 mL | 10 mL | 10 mL | 10 mL | 10 mL | 10 mL | 10 mL |
E. coli Painting (3)
We draw various pictures with ink brush.
- Bacteria which used as color
ptet-GFP, ptet-RFP, ptet-CFP
23:45-
We cultured painted plate at 37 degrees Celsius.
