Team:Chiba/Notebook/Calendar/14 September 2009

From 2009.igem.org

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([[Team:Chiba/Notebook/Calendar/13_September_2009|13_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/15_September_2009|15_September_2009]])
([[Team:Chiba/Notebook/Calendar/13_September_2009|13_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/15_September_2009|15_September_2009]])
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== Check of Sender's AHL generation (3) ==
 +
 +
 +
*Main culture
 +
8:15-
 +
 +
 +
*Measuring OD
 +
13:20
 +
 +
OD<sub>600</sub>=1.15
 +
 +
 +
14:00
 +
 +
OD<sub>600</sub>=1.65
 +
 +
 +
14:35
 +
 +
OD<sub>600</sub>=2.13
 +
 +
 +
*Wash
 +
14:40
 +
 +
First, we dispensed culture solution about 12.5 mL to each 10 centrifuging tubes.
 +
 +
 +
14:55
 +
 +
Next, we centrifuged samples 3 times(rpm=3000, 5 min, 20 degrees Celsius).
 +
 +
 +
*Centrifugation
 +
16:05
 +
 +
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.--
 +
 +
 +
19:15
 +
 +
We prepared mixture and made solidified media using this.
 +
 +
mixture : 10 mL LB-agar solidified medium and 10 mL supernatant solution
 +
 +
 +
19:30
 +
 +
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 ===
 +
==== 19:30 Start ====
 +
<gallery>
 +
Image:Chiba_1409_1930_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_1930_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_1930_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_1930_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_1930_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_1930_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_1930_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_1930_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_1930_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_1930_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_1930_NC.jpg|N.C
 +
</gallery>
 +
 +
==== 20:15 ====
 +
<gallery>
 +
Image:Chiba_1409_2015_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_2015_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_2015_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2015_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2015_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2015_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2015_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2015_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2015_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2015_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2015_NC.jpg|N.C
 +
</gallery>
 +
 +
==== 20:30 ====
 +
<gallery>
 +
Image:Chiba_1409_2030_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_2030_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_2030_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2030_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2030_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2030_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2030_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2030_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2030_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2030_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2030_NC.jpg|N.C
 +
</gallery>
 +
 +
==== 21:00 ====
 +
<gallery>
 +
Image:Chiba_1409_2100_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_2100_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_2100_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2100_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2100_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2100_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2100_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2100_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2100_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2100_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2100_NC.jpg|N.C
 +
</gallery>
 +
 +
 +
==== 21:30 ====
 +
<gallery>
 +
Image:Chiba_1409_2130_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_2130_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_2130_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2130_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2130_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2130_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2130_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2130_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2130_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2130_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2130_NC.jpg|N.C
 +
</gallery>
 +
 +
==== 22:00 ====
 +
<gallery>
 +
Image:Chiba_1409_2200_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_2200_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_2200_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2200_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2200_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2200_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2200_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2200_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2200_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2200_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2200_NC.jpg|N.C
 +
</gallery>
 +
 +
==== 22:30 ====
 +
<gallery>
 +
Image:Chiba_1409_2230_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_2230_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_2230_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2230_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2230_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2230_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2230_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2230_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2230_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2230_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2230_NC.jpg|N.C
 +
</gallery>
 +
 +
==== 23:00 ====
 +
<gallery>
 +
Image:Chiba_1409_2300_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_2300_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_2300_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2300_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2300_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2300_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2300_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2300_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2300_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2300_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2300_NC.jpg|N.C
 +
</gallery>
 +
 +
==== 23:45 ====
 +
<gallery>
 +
Image:Chiba_1409_2345_1.jpg|(1)AHL 0 min.
 +
Image:Chiba_1409_2345_2.jpg|(2)AHL 15 min.
 +
Image:Chiba_1409_2345_3.jpg|(3) 30 min.
 +
Image:Chiba_1409_2345_4.jpg|(4) 45 min.
 +
Image:Chiba_1409_2345_5.jpg|(5) 60 min.
 +
Image:Chiba_1409_2345_6.jpg|(6) 75 min.
 +
Image:Chiba_1409_2345_7.jpg|(7) 90 min.
 +
Image:Chiba_1409_2345_8.jpg|(8) 105 min.
 +
Image:Chiba_1409_2345_9.jpg|(9) 120 min.
 +
Image:Chiba_1409_2345_10.jpg|(10) 135 min.
 +
Image:Chiba_1409_2345_NC.jpg|N.C
 +
</gallery>
 +
 +
== スタブからぷらすみどとる。 ==
 +
 +
*Prior culture
 +
 +
22:00
 +
 +
We did prior culture using glycerol stock.
 +
 +
(1)[http://partsregistry.org/Part:BBa_K091118 BBa_K091118]
 +
 +
(2)[http://partsregistry.org/Part:BBa_K091134 BBa_K091134]
 +
 +
(3)[http://partsregistry.org/Part:BBa_S03956 BBa_S03956]
 +
 +
(4)[http://partsregistry.org/Part:BBa_I731012 BBa_I731012]
 +
 +
 +
 +
Continuation is [https://2009.igem.org/Team:Chiba/Notebook/Calendar/15_September_2009 here].
 +
 +
 +
== Lab Notebook ==
 +
 +
[[Image:Chiba-Labwork-14Sep09.jpg|200px]] [[Image:Chiba-Labwork-14Sep09-2.jpg|200px]]  [[Image:Chiba-Labwork-14Sep09-3.jpg|200px]]

Latest revision as of 03:08, 23 September 2009

>Go to the Notebook page

(13_September_2009 <|>15_September_2009)


Contents

Check of Sender's AHL generation (3)

  • Main culture

8:15-


  • Measuring OD

13:20

OD600=1.15


14:00

OD600=1.65


14:35

OD600=2.13


  • Wash

14:40

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


14:55

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


  • Centrifugation

16:05

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.--


19:15

We prepared mixture and made solidified media using this.

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


19:30

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

19:30 Start

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C

20:15

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C

20:30

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C

21:00

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C


21:30

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C

22:00

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C

22:30

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C

23:00

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C

23:45

(1)AHL 0 min.

(2)AHL 15 min.

(3) 30 min.

(4) 45 min.

(5) 60 min.

(6) 75 min.

(7) 90 min.

(8) 105 min.

(9) 120 min.

(10) 135 min.

N.C

スタブからぷらすみどとる。

  • Prior culture

22:00

We did prior culture using glycerol stock.

(1)[http://partsregistry.org/Part:BBa_K091118 BBa_K091118]

(2)[http://partsregistry.org/Part:BBa_K091134 BBa_K091134]

(3)[http://partsregistry.org/Part:BBa_S03956 BBa_S03956]

(4)[http://partsregistry.org/Part:BBa_I731012 BBa_I731012]


Continuation is here.


Lab Notebook

Chiba-Labwork-14Sep09.jpg Chiba-Labwork-14Sep09-2.jpg Chiba-Labwork-14Sep09-3.jpg

Retrieved from "http://2009.igem.org/Team:Chiba/Notebook/Calendar/14_September_2009"