Team:Chiba/Notebook/Calendar/22 September 2009
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([[Team:Chiba/Notebook/Calendar/21_September_2009|21_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/23_September_2009|23_September_2009]]) | ([[Team:Chiba/Notebook/Calendar/21_September_2009|21_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/23_September_2009|23_September_2009]]) | ||
+ | |||
+ | |||
+ | |||
+ | == Transformation(1)-2 == | ||
+ | Yesterday's operation is [https://2009.igem.org/Team:Chiba/Notebook/Calendar/21_September_2009#Transformation.281.29-1 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 | ||
+ | |||
+ | [http://partsregistry.org/Part:BBa_T9002 BBa_T9002] | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | == To judge character of LuxR mutants (1)-2 == | ||
+ | Yesterday's operation is [https://2009.igem.org/Team:Chiba/Notebook/Calendar/21_September_2009 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 | ||
+ | |||
+ | <table width="300" border="1" cellpadding="0" cellspacing="0" bordercolor="#000000"><tr> | ||
+ | <td width="150">Wild Type x 3 well</td> | ||
+ | <td width="150">Mutant 8 x 3 well</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Mutant 1</td> | ||
+ | <td>Mutant 9</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Mutant 2</td> | ||
+ | <td>Mutant 10</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Mutant 3</td> | ||
+ | <td>Mutant 11</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Mutant 4</td> | ||
+ | <td>Mutant L1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Mutant 5</td> | ||
+ | <td>Mutant L2</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Mutant 6</td> | ||
+ | <td>Negative Control</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Mutant 7</td> | ||
+ | <td>(Nothing)</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | '''Pictures are here.''' | ||
+ | |||
+ | === observed condition of fluorescence every 30 min === | ||
+ | ==== 22:35 Start ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2235_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2235_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2235_1000.jpg|(3 )AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | ==== 23:05 ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2305_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2305_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2305_1000.jpg|(3 )AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | ==== 23:35 ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2335_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2335_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2335_1000.jpg|(3 )AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | ==== 24:05 ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2405_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2405_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2405_1000.jpg|(3 )AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | ==== 24:35 ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2435_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2435_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2435_1000.jpg|(3 )AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | ==== 25:05 ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2505_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2505_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2505_1000.jpg|(3 )AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | ==== 25:35 ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2535_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2535_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2535_1000.jpg|(3 )AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | ==== 26:05 ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2605_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2605_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2605_1000.jpg|(3)AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | ==== 26:35 ==== | ||
+ | <gallery> | ||
+ | Image:Chiba_2209_2635_0.jpg|(1) AHL 0 nM | ||
+ | Image:Chiba_2209_2635_10.jpg|(2) AHL 10 nM | ||
+ | Image:Chiba_2209_2635_1000.jpg|(3 )AHL 1000nM | ||
+ | </gallery> | ||
+ | |||
+ | == ''E''. coli Painting (2) == | ||
+ | We painted some pictures using excess culture(LuxR Mutant 1) and cultured these. | ||
+ | |||
+ | |||
+ | |||
+ | '''Pictures are [https://2009.igem.org/wiki/index.php?title=Team:Chiba/Notebook/Calendar/23_September_2009 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). |
Latest revision as of 17:12, 25 September 2009
(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
[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]
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
23:05
23:35
24:05
24:35
25:05
25:35
26:05
26:35
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).