Team:Chiba/Notebook/Calendar/22 September 2009

From 2009.igem.org

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([[Team:Chiba/Notebook/Calendar/21_September_2009|21_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/23_September_2009|23_September_2009]])
([[Team:Chiba/Notebook/Calendar/21_September_2009|21_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/23_September_2009|23_September_2009]])
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== Transformation(1)-2 ==
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Yesterday's operation is [https://2009.igem.org/Team:Chiba/Notebook/Calendar/21_September_2009#Transformation.281.29-1 here].
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*Today's operation
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We count number of colonies of each plate.
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11:30
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コロニーをつついて培養
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pCIA3-LuxR
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LacZ alpha protein generator
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23:50
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mCherryのコロニーができたのでつついて培養
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グリストをつついて培養
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LuxI
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[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]
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Revision as of 03:27, 25 September 2009

>Go to the Notebook page

(21_September_2009 <|>23_September_2009)


Contents

Transformation(1)-2

Yesterday's operation is here.


  • Today's operation

We count number of colonies of each plate.


11:30

コロニーをつついて培養

pCIA3-LuxR

LacZ alpha protein generator


23:50

mCherryのコロニーができたのでつついて培養

グリストをつついて培養

LuxI

[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]



To judge character of LuxR mutants (1)-2

Yesterday's operation is here.


  • Today's operation

11:00-

We transplanted E.coli by 48 pins to NC filter and cultured it.


22:30

We transplanted E.coli, which has been cultured on NC filter, to solid medium which contains each concentration of AHL.

AHL concentration is : 0, 10, and 1000 nM


We decided this time is T=0 and observed condition of fluorescence every 30 min.


  • Mutants' Location
Wild Type x 3 well Mutant 8 x 3 well
Mutant 1 Mutant 9
Mutant 2 Mutant 10
Mutant 3 Mutant 11
Mutant 4 Mutant L1
Mutant 5 Mutant L2
Mutant 6 Negative Control
Mutant 7 (Nothing)


Pictures are here.

observed condition of fluorescence every 30 min

22:35 Start

(1) AHL 0 nM

(2) AHL 10 nM

(3 )AHL 1000nM

23:05

(1) AHL 0 nM

(2) AHL 10 nM

(3 )AHL 1000nM

23:35

(1) AHL 0 nM

(2) AHL 10 nM

(3 )AHL 1000nM

24:05

(1) AHL 0 nM

(2) AHL 10 nM

(3 )AHL 1000nM

24:35

(1) AHL 0 nM

(2) AHL 10 nM

(3 )AHL 1000nM

25:05

(1) AHL 0 nM

(2) AHL 10 nM

(3 )AHL 1000nM

25:35

(1) AHL 0 nM

(2) AHL 10 nM

(3 )AHL 1000nM

26:05

(1) AHL 0 nM

(2) AHL 10 nM

(3)AHL 1000nM

26:35

(1) AHL 0 nM

(2) AHL 10 nM

(3 )AHL 1000nM

E. coli Painting (2)

To judge character of LuxR mutants (2)-1

We poured 1 mL of LB-Amp, Cm liquid medium in 96 deep well and added glycerol stocks.


11:45-

We cultured and shook it at 37 degrees Celsius.


23:20-

We transplanted E.coli by 48 pins to NC filter and cultured it.

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