Team:Chiba/Notebook/Calendar/26 September 2009

From 2009.igem.org

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== E.coli Painting(3) ==
 +
<gallery>
 +
Image:Chiba_DSCF0478.jpg|RFP, GFP. CFP
 +
</gallery>
== Examine limit of AHL generation(2)-2 ==
== Examine limit of AHL generation(2)-2 ==
Line 13: Line 18:
*Today's operation
*Today's operation
11:35-
11:35-
 +
 +
We transplanted E.coli to NC filter and put these on each plates.
 +
 +
And then we took pictures every 30 min.
 +
 +
 +
'''Picture are here.'''
 +
 +
 +
== Pictures ==
 +
=== 11:35 ===
 +
<gallery>
 +
image:Chiba_DSCF0502.jpg|Negative Control
 +
Image:Chiba_DSCF0489.jpg|1
 +
Image:Chiba_DSCF0490.jpg|2
 +
Image:Chiba_DSCF0491.jpg|3
 +
Image:Chiba_DSCF0492.jpg|8
 +
Image:Chiba_DSCF0493.jpg|4
 +
Image:Chiba_DSCF0494.jpg|5
 +
Image:Chiba_DSCF0495.jpg|6
 +
Image:Chiba_DSCF0496.jpg|7
 +
</gallery>
 +
 +
 +
=== 12:05 ===
 +
<gallery>
 +
image:Chiba_DSCF0503.jpg|Negative Control
 +
Image:Chiba_DSCF0504.jpg|1
 +
Image:Chiba_DSCF0505.jpg|2
 +
Image:Chiba_DSCF0506.jpg|3
 +
Image:Chiba_DSCF0507.jpg|8
 +
Image:Chiba_DSCF0508.jpg|4
 +
Image:Chiba_DSCF0509.jpg|5
 +
Image:Chiba_DSCF0510.jpg|6
 +
Image:Chiba_DSCF0511.jpg|7
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</gallery>
 +
 +
 +
 +
=== 12:35 ===
 +
<gallery>
 +
image:Chiba_DSCF0512.jpg|Negative Control
 +
Image:Chiba_DSCF0513.jpg|1
 +
Image:Chiba_DSCF0514.jpg|2
 +
Image:Chiba_DSCF0515.jpg|3
 +
Image:Chiba_DSCF0516.jpg|8
 +
Image:Chiba_DSCF0517.jpg|4
 +
Image:Chiba_DSCF0518.jpg|5
 +
Image:Chiba_DSCF0519.jpg|6
 +
Image:Chiba_DSCF0520.jpg|7
 +
</gallery>
 +
 +
 +
 +
=== 13:05 ===
 +
<gallery>
 +
image:Chiba_DSCF0521.jpg|Negative Control
 +
Image:Chiba_DSCF0522.jpg|1
 +
Image:Chiba_DSCF0523.jpg|2
 +
Image:Chiba_DSCF0524.jpg|3
 +
Image:Chiba_DSCF0525.jpg|8
 +
Image:Chiba_DSCF0526.jpg|4
 +
Image:Chiba_DSCF0527.jpg|5
 +
Image:Chiba_DSCF0528.jpg|6
 +
Image:Chiba_DSCF0529.jpg|7
 +
</gallery>
 +
 +
 +
 +
=== 13:35 ===
 +
<gallery>
 +
image:Chiba_DSCF0530.jpg|Negative Control
 +
Image:Chiba_DSCF0531.jpg|1
 +
Image:Chiba_DSCF0532.jpg|2
 +
Image:Chiba_DSCF0533.jpg|3
 +
Image:Chiba_DSCF0534.jpg|8
 +
Image:Chiba_DSCF0535.jpg|4
 +
Image:Chiba_DSCF0536.jpg|5
 +
Image:Chiba_DSCF0537.jpg|6
 +
Image:Chiba_DSCF0538.jpg|7
 +
</gallery>
 +
 +
 +
 +
=== 14:05 ===
 +
<gallery>
 +
image:Chiba_DSCF0539.jpg|Negative Control
 +
Image:Chiba_DSCF0540.jpg|1
 +
Image:Chiba_DSCF0541.jpg|2
 +
Image:Chiba_DSCF0542.jpg|3
 +
Image:Chiba_DSCF0543.jpg|8
 +
Image:Chiba_DSCF0544.jpg|4
 +
Image:Chiba_DSCF0545.jpg|5
 +
Image:Chiba_DSCF0546.jpg|6
 +
Image:Chiba_DSCF0547.jpg|7
 +
</gallery>
 +
 +
 +
 +
=== 14:35 ===
 +
<gallery>
 +
image:Chiba_DSCF0548.jpg|Negative Control
 +
Image:Chiba_DSCF0550.jpg|1
 +
Image:Chiba_DSCF0551.jpg|2
 +
Image:Chiba_DSCF0552.jpg|3
 +
Image:Chiba_DSCF0553.jpg|8
 +
Image:Chiba_DSCF0554.jpg|4
 +
Image:Chiba_DSCF0555.jpg|5
 +
Image:Chiba_DSCF0556.jpg|6
 +
Image:Chiba_DSCF0557.jpg|7
 +
</gallery>
 +
 +
 +
 +
=== 15:05 ===
 +
<gallery>
 +
image:Chiba_DSCF0558.jpg|Negative Control
 +
Image:Chiba_DSCF0559.jpg|1
 +
Image:Chiba_DSCF0560.jpg|2
 +
Image:Chiba_DSCF0562.jpg|3
 +
Image:Chiba_DSCF0563.jpg|8
 +
Image:Chiba_DSCF0564.jpg|4
 +
Image:Chiba_DSCF0565.jpg|5
 +
Image:Chiba_DSCF0566.jpg|6
 +
Image:Chiba_DSCF0568.jpg|7
 +
</gallery>
 +
 +
== Making colors ==
 +
=== PCR ===
 +
*Template
 +
1 : ECFP([http://partsregistry.org/Part:BBa_I6057 BBa_I6057])
 +
 +
2 : mCherry([http://partsregistry.org/Part:BBa_E2060 BBa_E2060])
 +
 +
3 : V-YFP([http://partsregistry.org/Part:BBa_K084003 BBa_K084003])
 +
 +
4 : mOrange([http://partsregistry.org/Part:BBa_E2050 BBa_E2050])
 +
 +
5 : LacZ &alpha;([http://partsregistry.org/Part:BBa_T9003 BBa_T9003])
 +
 +
6 : mRFP without Nco1 site
 +
 +
 +
 +
14:45
 +
 +
We drew up mixtures for PCR.
 +
 +
 +
*Element of mixtures
 +
 +
<table width="200" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
<td width="100">Template</td>
 +
<td width="100">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Primer(F)</td>
 +
<td>2.5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Primer(R)</td>
 +
<td>2.5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer(x 10)</td>
 +
<td>5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>dNTP</td>
 +
<td>4 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>DNA pol.</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>34 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Total</td>
 +
<td>50 &mu;L</td>
 +
</tr>
 +
</table>
 +
 +
 +
15:20-
 +
 +
PCR
 +
 +
=== Gel electrophoresis ===
 +
15:30
 +
 +
We made gel for electrophoresis.
 +
 +
agar 0.14 g
 +
 +
TAE  20 mL
 +
 +
 +
17:25-
 +
 +
Gel electrophoresis(135 V, 27 min)

Latest revision as of 05:56, 27 September 2009

>Go to the Notebook page

(25_September_2009 <|>27_September_2009)


Contents

E.coli Painting(3)

RFP, GFP. CFP

Examine limit of AHL generation(2)-2

Yesterday's operation is here.


  • Today's operation

11:35-

We transplanted E.coli to NC filter and put these on each plates.

And then we took pictures every 30 min.


Picture are here.


Pictures

11:35

Negative Control

1

2

3

8

4

5

6

7


12:05

Negative Control

1

2

3

8

4

5

6

7


12:35

Negative Control

1

2

3

8

4

5

6

7


13:05

Negative Control

1

2

3

8

4

5

6

7


13:35

Negative Control

1

2

3

8

4

5

6

7


14:05

Negative Control

1

2

3

8

4

5

6

7


14:35

Negative Control

1

2

3

8

4

5

6

7


15:05

Negative Control

1

2

3

8

4

5

6

7

Making colors

PCR

  • Template

1 : ECFP([http://partsregistry.org/Part:BBa_I6057 BBa_I6057])

2 : mCherry([http://partsregistry.org/Part:BBa_E2060 BBa_E2060])

3 : V-YFP([http://partsregistry.org/Part:BBa_K084003 BBa_K084003])

4 : mOrange([http://partsregistry.org/Part:BBa_E2050 BBa_E2050])

5 : LacZ α([http://partsregistry.org/Part:BBa_T9003 BBa_T9003])

6 : mRFP without Nco1 site


14:45

We drew up mixtures for PCR.


  • Element of mixtures
Template 1 μL
Primer(F) 2.5 μL
Primer(R) 2.5 μL
Buffer(x 10) 5 μL
dNTP 4 μL
DNA pol. 1 μL
dH2O 34 μL
Total 50 μL


15:20-

PCR

Gel electrophoresis

15:30

We made gel for electrophoresis.

agar 0.14 g

TAE 20 mL


17:25-

Gel electrophoresis(135 V, 27 min)

Retrieved from "http://2009.igem.org/Team:Chiba/Notebook/Calendar/26_September_2009"