Team:Chiba/Notebook/Calendar/25 September 2009

From 2009.igem.org

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<table width="300" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
<table width="300" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
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<td width="150">Mutant 1 x 3 well</td>
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<td width="150">Wild Type x 3 well</td>
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<td width="150">Mutant 9 x 3 well</td>
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<td width="150">Mutant 8 x 3 well</td>
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<td>Mutant 1</td>
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<td>Mutant 9</td>
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<td>Mutant 6</td>
<td>Mutant 6</td>
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<td>Wild Type</td>
 
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<td>Mutant 7</td>
 
<td>Negative Control</td>
<td>Negative Control</td>
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<td>Mutant 8</td>
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<td>Mutant 7</td>
<td>(Nothing)</td>
<td>(Nothing)</td>
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'''Picture are here.'''
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'''Pictures are here.'''
==== 22:20 Start ====
==== 22:20 Start ====
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We cultured painted plate at 37 degrees Celsius.
We cultured painted plate at 37 degrees Celsius.
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'''Picture is [https://2009.igem.org/Team:Chiba/Notebook/Calendar/26_September_2009 here].'''

Latest revision as of 08:04, 28 September 2009

>Go to the Notebook page

(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

(1) AHL 0 nM

(2) AHL 1 nM

(3) AHL 10 nM

(4)AHL 100nM

(5)AHL 1000nM

23:20

(1) AHL 0 nM

(2) AHL 1 nM

(3) AHL 10 nM

(4)AHL 100nM

(5)AHL 1000nM

23:50

(1) AHL 0 nM

(2) AHL 1 nM

(3) AHL 10 nM

(4)AHL 100nM

(5)AHL 1000nM

24:20

(1) AHL 0 nM

(2) AHL 1 nM

(3) AHL 10 nM

(4)AHL 100nM

(5)AHL 1000nM

24:50

(1) AHL 0 nM

(2) AHL 1 nM

(3) AHL 10 nM

(4)AHL 100nM

(5)AHL 1000nM

25:20

(1) AHL 0 nM

(2) AHL 1 nM

(3) AHL 10 nM

(4)AHL 100nM

(5)AHL 1000nM

25:50

(1) AHL 0 nM

(2) AHL 1 nM

(3) AHL 10 nM

(4)AHL 100nM

(5)AHL 1000nM

26:20

(1) AHL 0 nM

(2) AHL 1 nM

(3) AHL 10 nM

(4)AHL 100nM

(5)AHL 1000nM

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.

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