Team:Chiba/Notebook/Calendar/22 September 2009

From 2009.igem.org

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== LuxR Mutantの個性決める実験の写真だけうｐ ==
+
 
 +
== 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>
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<td width="150">Mutant 8 x 3 well</td>
 +
</tr>
 +
<tr>
 +
<td>Mutant 1</td>
 +
<td>Mutant 9</td>
 +
</tr>
 +
<tr>
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<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>
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<td>(Nothing)</td>
 +
</tr>
 +
</table>
 +
 
 +
 
 +
'''Pictures are here.'''
 +
 
 +
=== observed condition of fluorescence every 30 min ===
==== 22:35 Start ====
==== 22:35 Start ====
<gallery>
<gallery>
Line 58: Line 154:
==== 26:05 ====
==== 26:05 ====
<gallery>
<gallery>
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Image:Chiba_2209_2205_0.jpg|(1) AHL 0 nM
+
Image:Chiba_2209_2605_0.jpg|(1) AHL 0 nM
-
Image:Chiba_2209_2205_10.jpg|(2) AHL 10 nM
+
Image:Chiba_2209_2605_10.jpg|(2) AHL 10 nM
-
Image:Chiba_2209_2205_1000.jpg|(3 )AHL 1000nM
+
Image:Chiba_2209_2605_1000.jpg|(3)AHL 1000nM
</gallery>
</gallery>
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Image:Chiba_2209_2635_1000.jpg|(3 )AHL 1000nM
Image:Chiba_2209_2635_1000.jpg|(3 )AHL 1000nM
</gallery>
</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

>Go to the Notebook page

(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

(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).

Retrieved from "http://2009.igem.org/Team:Chiba/Notebook/Calendar/22_September_2009"