Team:Chiba/Notebook/Calendar/23 September 2009

From 2009.igem.org

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(LuxR Mutantの個性決める実験の写真だけうp)
(To judge character of LuxR mutants (2)-2)
 
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Latest revision as of 08:05, 28 September 2009

>Go to the Notebook page

(22_September_2009 <|>24_September_2009)

Contents

E. coli Painting (2)

We draw various pictures with ink brush and throwaway chopsticks.

Yesterday's operation is here.

To judge character of LuxR mutants (2)-2

Yesterday's operation is here.


  • Today's operation

10:30

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

AHL concentration is : 0, 1, 10, 100, and 1000 nM


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


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


Pictures are here.

photo

10:30 Start

11:00

11:30

12:00

12:30

13:00

13:30

14:00

14:30

Examine limit of AHL generation(1)-2

Yesterday's operation is here.


  • Today's operation

11:00

We did main culture.


22:00

We measured OD.

OD600=1.98


Then, we spun down this culture solution and add 12.5 mL of flesh LB-Amp.


22:10

We add 12.5 μL of IPTG to the culture solution and started culture at 37 degrees Celsius.

Transformation(2)-1

  • Plasmid

plux-GFP(Cm, p15A)


  • Competent Cells

JW1262(pCIA3-LuxR(Amp))

JW1262(Null)


21:35

We transplanted these bacteria each plates.

pCIA3-LuxR and plux-GFP --> LB-Amp, Cm plate

plux-GFP only --> LB-Cm plate


21:50-

We cultured these plates.