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) ==
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*Main culture
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8:15-
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*Measuring OD
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13:20
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OD<sub>600</sub>=1.15
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14:00
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OD<sub>600</sub>=1.65
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14:35
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OD<sub>600</sub>=2.13
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*Wash
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14:40
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First, we dispensed culture solution about 12.5 mL to each 10 centrifuging tubes.
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14:55
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Next, we centrifuged samples 3 times(rpm=3000, 5 min, 20 degrees Celsius).
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*Centrifugation
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16:05
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We added more 12.5 mL of liquid medium and 12.5 μL of 0.1 M IPTG to each tubes.
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After that, we centrifuged one of samples, and cultured the others at 30 degrees Celsius.
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And then, we picked 10 mL of supernatant solution and saved it.
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-- We picked supernatant solution every 15 min.--
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19:15
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We prepared mixture and made solidified media using this.
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mixture : 10 mL LB-agar solidified medium and 10 mL supernatant solution
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19:30
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We put NC filters on plates which has been prepared a little while ago.
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After that we started observation of GFP fluorescence.
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'''Pictures are here.'''

Revision as of 05:40, 15 September 2009

>Go to the Notebook page

(13_September_2009 <|>15_September_2009)


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.

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