Team:Chiba/Notebook/Calendar/23 September 2009

From 2009.igem.org

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([[Team:Chiba/Notebook/Calendar/22_September_2009|22_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/24_September_2009|24_September_2009]])
([[Team:Chiba/Notebook/Calendar/22_September_2009|22_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/24_September_2009|24_September_2009]])
-
== ''E''. coli Painting ==
+
 
 +
== ''E''. coli Painting (2) ==
 +
We draw various pictures with ink brush and throwaway chopsticks.
 +
 
 +
Yesterday's operation is [https://2009.igem.org/wiki/index.php?title=Team:Chiba/Notebook/Calendar/22_September_2009 here].
 +
 
<gallery>
<gallery>
Image:Chiba_2309_paint_0.jpg|2009 iGEM Chiba  
Image:Chiba_2309_paint_0.jpg|2009 iGEM Chiba  
Line 11: Line 16:
Image:Chiba_2309_paint_3.jpg|pugyaaa
Image:Chiba_2309_paint_3.jpg|pugyaaa
Image:Chiba_2309_paint_4.jpg|aiueo
Image:Chiba_2309_paint_4.jpg|aiueo
-
Image:Chiba_2309_paint_5.jpg|
+
Image:Chiba_2309_paint_5.jpg|using throwaway chopsticks
</gallery>
</gallery>
-
== LuxR Mutantの個性決める実験の写真だけうｐ ==
+
== To judge character of LuxR mutants (2)-2 ==
 +
Yesterday's operation is [https://2009.igem.org/Team:Chiba/Notebook/Calendar/22_September_2009 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
 +
 
 +
<table width="300" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
<td width="150">Mutant 1 x 3 well</td>
 +
<td width="150">Mutant 9 x 3 well</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>Wild Type</td>
 +
</tr>
 +
<tr>
 +
<td>Mutant 7</td>
 +
<td>Negative Control</td>
 +
</tr>
 +
<tr>
 +
<td>Mutant 8</td>
 +
<td>(Nothing)</td>
 +
</tr>
 +
</table>
 +
 
 +
 
 +
'''Pictures are here.'''
 +
 
 +
=== photo ===
==== 10:30 Start ====
==== 10:30 Start ====
<gallery>
<gallery>
Line 36: Line 95:
<gallery>
<gallery>
Image:Chiba_2309_1130_0.jpg|(1) AHL 0 nM
Image:Chiba_2309_1130_0.jpg|(1) AHL 0 nM
-
Image:Chiba_2309_1130_10.jpg|(2) AHL 1 nM
+
Image:Chiba_2309_1130_1.jpg|(2) AHL 1 nM
Image:Chiba_2309_1130_10.jpg|(3) AHL 10 nM
Image:Chiba_2309_1130_10.jpg|(3) AHL 10 nM
-
Image:Chiba_2309_1130_1000.jpg|(4)AHL 100nM
+
Image:Chiba_2309_1130_100.jpg|(4)AHL 100nM
Image:Chiba_2309_1130_1000.jpg|(5)AHL 1000nM
Image:Chiba_2309_1130_1000.jpg|(5)AHL 1000nM
</gallery>
</gallery>
Line 54: Line 113:
<gallery>
<gallery>
Image:Chiba_2309_1230_0.jpg|(1) AHL 0 nM
Image:Chiba_2309_1230_0.jpg|(1) AHL 0 nM
-
Image:Chiba_2309_1230_10.jpg|(2) AHL 1 nM
+
Image:Chiba_2309_1230_1.jpg|(2) AHL 1 nM
Image:Chiba_2309_1230_10.jpg|(3) AHL 10 nM
Image:Chiba_2309_1230_10.jpg|(3) AHL 10 nM
-
Image:Chiba_2309_1230_1000.jpg|(4)AHL 100nM
+
Image:Chiba_2309_1230_100.jpg|(4)AHL 100nM
Image:Chiba_2309_1230_1000.jpg|(5)AHL 1000nM
Image:Chiba_2309_1230_1000.jpg|(5)AHL 1000nM
</gallery>
</gallery>
Line 72: Line 131:
<gallery>
<gallery>
Image:Chiba_2309_1330_0.jpg|(1) AHL 0 nM
Image:Chiba_2309_1330_0.jpg|(1) AHL 0 nM
-
Image:Chiba_2309_1330_10.jpg|(2) AHL 1 nM
+
Image:Chiba_2309_1330_1.jpg|(2) AHL 1 nM
Image:Chiba_2309_1330_10.jpg|(3) AHL 10 nM
Image:Chiba_2309_1330_10.jpg|(3) AHL 10 nM
-
Image:Chiba_2309_1330_1000.jpg|(4)AHL 100nM
+
Image:Chiba_2309_1330_100.jpg|(4)AHL 100nM
Image:Chiba_2309_1330_1000.jpg|(5)AHL 1000nM
Image:Chiba_2309_1330_1000.jpg|(5)AHL 1000nM
</gallery>
</gallery>
Line 95: Line 154:
Image:Chiba_2309_1430_1000.jpg|(5)AHL 1000nM
Image:Chiba_2309_1430_1000.jpg|(5)AHL 1000nM
</gallery>
</gallery>
 +
 +
== Examine limit of AHL generation(1)-2 ==
 +
Yesterday's operation is [https://2009.igem.org/wiki/index.php?title=Team:Chiba/Notebook/Calendar/22_September_2009 here].
 +
 +
 +
*Today's operation
 +
11:00
 +
 +
We did main culture.
 +
 +
 +
 +
22:00
 +
 +
We measured OD.
 +
 +
OD<sub>600</sub>=1.98
 +
 +
 +
Then, we spun down this culture solution and add 12.5 mL of flesh LB-Amp.
 +
 +
 +
 +
22:10
 +
 +
We add 12.5 &mu;L 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.

Latest revision as of 08:05, 28 September 2009

>Go to the Notebook page

(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.

Retrieved from "http://2009.igem.org/Team:Chiba/Notebook/Calendar/23_September_2009"