Team:Chiba/Notebook/Calendar/13 September 2009

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<html><link rel="stylesheet" href="https://2008.igem.org/wiki/index.php?title=User:Maiko/chiba.css&action=raw&ctype=text/css" type="text/css" /></html>
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>[[Team:Chiba/Notebook|Go to the Notebook page]]
>[[Team:Chiba/Notebook|Go to the Notebook page]]
([[Team:Chiba/Notebook/Calendar/12_September_2009|12_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/14_September_2009|14_September_2009]])
([[Team:Chiba/Notebook/Calendar/12_September_2009|12_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/14_September_2009|14_September_2009]])
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 +
== Check of Sender's AHL generation (3) ==
 +
*Main culture
 +
7:10-
 +
 +
 +
*Measuring OD
 +
12:20
 +
 +
OD<sub>600</sub>=1.98
 +
 +
 +
*Wash
 +
12:40
 +
 +
First, we dispensed culture solution about 12.5 mL to each 10 centrifuging tubes.
 +
 +
 +
12:45
 +
 +
Next, we centrifuged samples 3 times(rpm=3000, 5 min, 20 degrees Celsius).
 +
 +
 +
 +
*Centrifugation
 +
13:50
 +
 +
We added more 12.5 mL of liquid medium and 12.5 &mu;L of 0.1 M IPTG to each tubes.
 +
 +
After that, we centrifuged one of samples, and cultured the others at 30 degrees Celsius.
 +
 +
And then, we picked 10 mL of supernatant solution and saved it.
 +
 +
 +
-- We picked supernatant solution every 15 min.--
 +
 +
 +
16:15
 +
 +
We prepared mixture and made solidified media using this.
 +
 +
:mixture : 10 mL LB-agar solidified medium and 10 mL supernatant solution
 +
 +
 +
 +
17:20
 +
 +
We transferred colonies onto NC filters and cultured it at 37 degrees Celsius.
 +
 +
 +
18:05
 +
 +
We put NC filters on plates which has been prepared a little while ago.
 +
 +
After that we started observation of GFP fluorescence.
 +
 +
 +
 +
'''Pictures are here.'''
 +
 +
 +
 +
=== Check of Sender's AHL generation ===
 +
==== 18:05 Start ====
 +
<gallery>
 +
Image:Chiba_1309_1805_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_1805_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_1805_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_1805_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_1805_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_1805_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_1805_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_1805_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_1805_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_1805_10.jpg|(10) 135 min.
 +
</gallery>
 +
 +
 +
==== 18:35 ====
 +
<gallery>
 +
Image:Chiba_1309_1835_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_1835_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_1835_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_1835_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_1835_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_1835_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_1835_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_1835_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_1835_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_1835_10.jpg|(10) 135 min.
 +
</gallery>
 +
 +
 +
==== 19:05 ====
 +
<gallery>
 +
Image:Chiba_1309_1905_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_1905_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_1905_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_1905_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_1905_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_1905_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_1905_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_1905_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_1905_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_1905_10.jpg|(10) 135 min.
 +
</gallery>
 +
 +
 +
 +
==== 19:35 ====
 +
<gallery>
 +
Image:Chiba_1309_1935_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_1935_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_1935_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_1935_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_1935_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_1935_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_1935_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_1935_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_1935_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_1935_10.jpg|(10) 135 min.
 +
Image:Chiba_1309_1935_NC.jpg|N.C.
 +
</gallery>
 +
 +
==== 20:05 ====
 +
<gallery>
 +
Image:Chiba_1309_2005_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_2005_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_2005_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_2005_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_2005_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_2005_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_2005_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_2005_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_2005_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_2005_10.jpg|(10) 135 min.
 +
Image:Chiba_1309_2005_NC.jpg|N.C.
 +
</gallery>
 +
 +
==== 20:35 ====
 +
<gallery>
 +
Image:Chiba_1309_2035_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_2035_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_2035_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_2035_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_2035_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_2035_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_2035_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_2035_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_2035_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_2035_10.jpg|(10) 135 min.
 +
</gallery>
 +
 +
==== 21:05 ====
 +
 +
(4)-(10) were became luminescent.
 +
 +
<gallery>
 +
Image:Chiba_1309_2105_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_2105_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_2105_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_2105_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_2105_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_2105_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_2105_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_2105_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_2105_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_2105_10.jpg|(10) 135 min.
 +
Image:Chiba_1309_2105_NC.jpg|N.C.
 +
</gallery>
 +
 +
 +
==== 21:35 ====
 +
 +
(3) has became luminescent.
 +
 +
<gallery>
 +
Image:Chiba_1309_2135_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_2135_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_2135_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_2135_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_2135_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_2135_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_2135_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_2135_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_2135_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_2135_10.jpg|(10) 135 min.
 +
Image:Chiba_1309_2135_NC.jpg|N.C.
 +
</gallery>
 +
 +
 +
==== 22:05 ====
 +
<gallery>
 +
Image:Chiba_1309_2205_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1309_2205_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1309_2205_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_2205_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_2205_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_2205_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_2205_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_2205_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_2205_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_2205_10.jpg|(10) 135 min.
 +
Image:Chiba_1309_2205_NC.jpg|N.C.
 +
</gallery>
 +
 +
==== 22:35 (14, Sep.)====
 +
<gallery>
 +
Image:Chiba_1409_2235_1.jpg|(1) 0 min.
 +
Image:Chiba_1409_2235_2.jpg|(2) 15 min.
 +
Image:Chiba_1409_2235_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2235_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2235_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2235_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2235_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2235_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2235_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2235_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2235_NC.jpg|N.C.
 +
</gallery>
 +
 +
 +
== Check of Sender's AHL generation (1) ==
 +
This Sample use the AHL, making by the Sender's culture without IPTG.
 +
 +
==== 20:00 ====
 +
<gallery>
 +
Image:Chiba_1309_2000_1.jpg|(1) 0 min.
 +
Image:Chiba_1309_2000_2.jpg|(2) 15 min.
 +
Image:Chiba_1309_2000_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_2000_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_2000_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_2000_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_2000_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_2000_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_2000_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_2000_10.jpg|(10) 135 min.
 +
Image:Chiba_1309_2000_NC.jpg|N.C.
 +
</gallery>
 +
 +
==== 20:30 ====
 +
<gallery>
 +
Image:Chiba_1309_2030_1.jpg|(1) 0 min.
 +
Image:Chiba_1309_2030_2.jpg|(2) 15 min.
 +
Image:Chiba_1309_2030_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_2030_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_2030_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_2030_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_2030_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_2030_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_2030_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_2030_10.jpg|(10) 135 min.
 +
Image:Chiba_1309_2030_NC.jpg|N.C.
 +
</gallery>
 +
 +
==== 22:35 (14, Sep.) ====
 +
<gallery>
 +
Image:Chiba_1309_2235_1.jpg|(1) 0 min.
 +
Image:Chiba_1309_2235_2.jpg|(2) 15 min.
 +
Image:Chiba_1309_2235_3.jpg|(3) 30 min.
 +
Image:Chiba_1309_2235_4.jpg|(4) 45 min.
 +
Image:Chiba_1309_2235_5.jpg|(5) 60 min.
 +
Image:Chiba_1309_2235_6.jpg|(6) 75 min.
 +
Image:Chiba_1309_2235_7.jpg|(7) 90 min.
 +
Image:Chiba_1309_2235_8.jpg|(8) 105 min.
 +
Image:Chiba_1309_2235_9.jpg|(9) 120 min.
 +
Image:Chiba_1309_2235_10.jpg|(10) 135 min.
 +
Image:Chiba_1309_2235_NC.jpg|N.C.
 +
</gallery>
 +
 +
== Handwritten Lab Notebook ==
 +
[[Image:Chiba-Labwork-12Sep09.jpg|200px]] [[Image:Chiba-Labwork-13Sep09.jpg|200px]]

Latest revision as of 18:33, 23 September 2009

>Go to the Notebook page

(12_September_2009 <|>14_September_2009)

Contents

Check of Sender's AHL generation (3)

  • Main culture

7:10-


  • Measuring OD

12:20

OD600=1.98


  • Wash

12:40

First, we dispensed culture solution about 12.5 mL to each 10 centrifuging tubes.


12:45

Next, we centrifuged samples 3 times(rpm=3000, 5 min, 20 degrees Celsius).


  • Centrifugation

13:50

We added more 12.5 mL of liquid medium and 12.5 μL of 0.1 M IPTG to each tubes.

After that, we centrifuged one of samples, and cultured the others at 30 degrees Celsius.

And then, we picked 10 mL of supernatant solution and saved it.


-- We picked supernatant solution every 15 min.--


16:15

We prepared mixture and made solidified media using this.

mixture : 10 mL LB-agar solidified medium and 10 mL supernatant solution


17:20

We transferred colonies onto NC filters and cultured it at 37 degrees Celsius.


18:05

We put NC filters on plates which has been prepared a little while ago.

After that we started observation of GFP fluorescence.


Pictures are here.


Check of Sender's AHL generation

18:05 Start


18:35


19:05


19:35

20:05

20:35

21:05

(4)-(10) were became luminescent.


21:35

(3) has became luminescent.


22:05

22:35 (14, Sep.)


Check of Sender's AHL generation (1)

This Sample use the AHL, making by the Sender's culture without IPTG.

20:00

20:30

22:35 (14, Sep.)

Handwritten Lab Notebook

Chiba-Labwork-12Sep09.jpg Chiba-Labwork-13Sep09.jpg