Team:Chiba/Notebook/Calendar/25 September 2009

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(24_September_2009 <|>26_September_2009)


Contents

To judge character of LuxR mutants (3)-2

Yesterday's operation is here.

  • Today's operation

10:45-

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


22:20 measurement start

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
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.

22:20 Start

23:20

23:50

24:20

24:50

25:20

25:50

26:20

Examine limit of AHL generation(2)-1

11:05-

We did prior culture.

glycerol stock : plux-GFP with pCIA3-LuxR


23:35-

We attenuated culture solution 1/105 and dispersed on plate.

Then we cultured this plate at 37 degrees Celsius.


And We made plates from each of mixture and 10 mL of LB-Amp, Cm solution medium.


  • Element of mixtures
Sample Namber 1 2 3 4 5 6 7 8
supernatant solution 5 mL --- --- --- --- --- --- 200 μL
1 --- 5 mL --- --- --- --- --- ---
2 --- --- 5 mL --- --- --- --- ---
3 --- --- --- 5 mL --- --- --- ---
4 --- --- --- --- 5 mL --- --- ---
5 --- --- --- --- --- 5 mL --- ---
6 --- --- --- --- --- --- 5 mL ---
LB-Amp,Cm(liquid) 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL 9.8 mL
Total 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL

E. coli Painting (3)

We draw various pictures with ink brush.


  • Bacteria which used as color

ptet-GFP, ptet-RFP, ptet-CFP


23:45-

We cultured painted plate at 37 degrees Celsius.


Picture is here.