"
Team:Chiba/Notebook/Calendar/22 September 2009
From 2009.igem.org
(21_September_2009 <|>23_September_2009)
Contents |
Transformation(1)-2
Yesterday's operation is here.
- Today's operation
We count number of colonies of each plate.
11:30
コロニーをつついて培養
pCIA3-LuxR
LacZ alpha protein generator
23:50
mCherryのコロニーができたのでつついて培養
グリストをつついて培養
LuxI
To judge character of LuxR mutants (1)-2
Yesterday's operation is here.
- Today's operation
11:00-
We transplanted E.coli by 48 pins to NC filter and cultured it.
22:30
We transplanted E.coli, which has been cultured on NC filter, to solid medium which contains each concentration of AHL.
AHL concentration is : 0, 10, and 1000 nM
We decided this time is T=0 and observed condition of fluorescence every 30 min.
- Mutants' Location
| Wild Type x 3 well | Mutant 8 x 3 well |
| Mutant 1 | Mutant 9 |
| Mutant 2 | Mutant 10 |
| Mutant 3 | Mutant 11 |
| Mutant 4 | Mutant L1 |
| Mutant 5 | Mutant L2 |
| Mutant 6 | Negative Control |
| Mutant 7 | (Nothing) |
Pictures are here.
observed condition of fluorescence every 30 min
22:35 Start
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3 )AHL 1000nM |
23:05
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3 )AHL 1000nM |
23:35
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3 )AHL 1000nM |
24:05
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3 )AHL 1000nM |
24:35
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3 )AHL 1000nM |
25:05
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3 )AHL 1000nM |
25:35
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3 )AHL 1000nM |
26:05
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3)AHL 1000nM |
26:35
 (1) AHL 0 nM |
 (2) AHL 10 nM |
 (3 )AHL 1000nM |
E. coli Painting (2)
We painted some pictures using excess culture(LuxR Mutant 1) and cultured these.
Pictures are here.
To judge character of LuxR mutants (2)-1
We poured 1 mL of LB-Amp, Cm liquid medium in 96 deep well and added glycerol stocks.
11:45-
We cultured and shook it at 37 degrees Celsius.
23:20-
We transplanted E.coli by 48 pins to NC filter and cultured it.
Examine limit of AHL generation(1)-1
23:00
We did prior culture(JW1908 glycerol stock, plac-LuxI, and 12.5 mL of LB-Amp).
