Team:Chiba/Notebook 3

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                                        <!-- CSS Tabs -->
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                                <!-- CSS Tabs -->
<li><a href="https://2010.igem.org/Team:Chiba"><span>Home</span></a></li>
<li><a href="https://2010.igem.org/Team:Chiba"><span>Home</span></a></li>
-
<li><a href="https://2010.igem.org/Team:Chiba/Team"><span>Team</span></a>
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<li><a href="https://2010.igem.org/Team:Chiba/Team"><span>Team</span></a></li>
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<ul style="z-index:1"> <br>
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<li><a href="https://2010.igem.org/Team:Chiba/Project"><span>Project</span></a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Team">Team</a></li> <li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Member">Member</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/School">School</a></li>
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</ul>
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</li>
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<li><a href="https://2010.igem.org/Team:Chiba/Project"><span>Project</span></a>
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<ul style="z-index:1"><br>
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                        <li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Project">Overall Project</a></li>
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<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Circuit_1">Circuit 1</a></li>
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-
<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Circuit_2">Circuit 2</a></li>
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-
</ul>
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-
</li>
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<li><a href="https://2010.igem.org/Team:Chiba/Parts"><span>Parts</span></a></li>
<li><a href="https://2010.igem.org/Team:Chiba/Parts"><span>Parts</span></a></li>
-
<li><a href="https://2010.igem.org/Team:Chiba/Modeling"><span>Modeling</span></a></li>
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<li><a href="https://2010.igem.org/Team:Chiba/Notebook"><span>Notebook</span></a></li>
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<li><a href="https://2010.igem.org/Team:Chiba/Result"><span>Result</span></a>
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-
<ul style="z-index:1"><br>
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-
                        <li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Plasmid1">Plasmid 1</a></li>
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-
<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Plasmid2">Plasmid 2</a></li>
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-
<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/lux_promoter">lux Promoter</a></li><br>
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-
</ul>
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-
</li>
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-
<li><a href="https://2010.igem.org/Team:Chiba/Notebook"><span>Notebook</span></a>
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-
<ul style="z-index:1"><br>
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-
                        <li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Notebook">Notebook 1</a></li>
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-
<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Notebook_2">Notebook 2</a></li>
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-
<li><a class="tablinks" href="https://2010.igem.org/Team:Chiba/Notebook_3">Notebook 3</a></li><br>
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-
</ul>
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-
</li>
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-
 
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<li><a href="https://2010.igem.org/Team:Chiba/Support"><span>Support</span></a></li>
<li><a href="https://2010.igem.org/Team:Chiba/Support"><span>Support</span></a></li>
-
<li><a href="https://2010.igem.org/Team:Chiba/Safety"><span>Safety</span></a></li>
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<li id="current"><a href="https://2010.igem.org/Team:Chiba/Safety"><span>Safety</span></a></li>
-
                                </ul>
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<!--- 2nd tab --->
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<!--- Background Contents End --->
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<!--- 2nd tab --->
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<!--- Main Contents Start --->
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<div id="search-controls">
-
<br>Lux inverter<br>
+
<form action="/Special:Search" id="searchform">
-
<html><h3>2010-10-8</h3></html>
+
<input id="searchInput" name="search" type="text" title="Search 2010.igem.org [f]" accesskey="f" value="" />
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<input type='submit' name="go" class="searchButton" id="searchGoButton" value="Go" title="Go to a page with this exact name if exists" />&nbsp;
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<style type="text/css">  
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<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
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<html xmlns="http://www.w3.org/1999/xhtml">
 +
        <head>
 +
                <meta http-equiv="Content-Type" content="text/html; charset=iso-8859-1" />
 +
                <title>Free CSS Navigation Menu Designs 2 at exploding-boy.com</title>
 +
                <style type="text/css">
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<!--
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img {
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border: none;
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}
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transformation
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/*- Menu Tabs 9--------------------------- */
-
BBa_R0061
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    #tabs9 {
-
BBa_E0240
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      float:left;
 +
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      padding:0;
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 +
      }
 +
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 +
      float:left;
 +
      background:url("tableft10.gif") no-repeat left top;
 +
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 +
      padding:0 0 0 2px;
 +
      text-decoration:none;
 +
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 +
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 +
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 +
      display:block;
 +
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 +
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 +
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                <div id="tabs9">
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                <ul>
 +
                                <!-- CSS Tabs -->
 +
<li><a href="https://2010.igem.org/Team:Chiba/Notebook"><span>Notebook 1</span></a></li>
 +
<li><a href="https://2010.igem.org/Team:Chiba/Notebook_2"><span>Notebook 2</span></a></li>
 +
<li><a href="https://2010.igem.org/Team:Chiba/Notebook_3"><span>Notebook 3</span></a></li>
 +
                        </ul>
 +
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<!--- Main Contents Start --->
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<br><br><br><br>
 +
<font size="5">Characterization of lux inverter</font><br><br>
 +
<html><h3>2010-10-8</h3></html>
 +
<pre>transformation
 +
 +
Plasmid:BBa_R0061
 +
        BBa_E0240
 +
strain:XL10-G
 +
 +
</pre>
<br>
<br>
<html><h3>2010-10-9</h3></html>
<html><h3>2010-10-9</h3></html>
-
Mini-prep
+
 
 +
<pre>Mini-prep (Sigma Aldrich)
 +
 
BBa_R0061
BBa_R0061
BBa_E0240
BBa_E0240
digestion
digestion
-
BBa_R0061    20 µL
 
-
10×Buffer    3 µL
 
-
SpeI　　　　　 1 µL
 
-
PstI　　　　　 1 µL
 
-
dH<sub>2</sub>O5 µL
 
--------------------
--------------------
 +
BBa_R0061      20 µL
 +
10×Buffer      3 µL
 +
SpeI　　　　　  1 µL
 +
PstI　　　　　  1 µL
 +
NFW            5 µL
 +
--------------------
 +
　　　　　　　 30 µL
--------------------
--------------------
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XbaI            1 µL
XbaI            1 µL
PstI　　　　　  1 µL
PstI　　　　　  1 µL
-
dH<sub>2</sub>O 5 µL
+
NFW            5 µL
--------------------
--------------------
-
 
+
              30 µL
 +
↓
gel extraction
gel extraction
-
<br>
+
</pre>
 +
 
<html><h3>2010-10-11</h3></html>
<html><h3>2010-10-11</h3></html>
-
 
+
<pre>
Ligation
Ligation
-
 
+
-----------------------
BBa_R0061 (70 ng)  1 µL       
BBa_R0061 (70 ng)  1 µL       
BBa_E0240 (100 ng) 2 µL
BBa_E0240 (100 ng) 2 µL
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10×Buffer        1 µL
10×Buffer        1 µL
T4 DNA Ligase      1 µL
T4 DNA Ligase      1 µL
-
dH<sub>2</sub>O    4 µL
+
NFW                4 µL
-
 
+
-----------------------
 +
                  10 µL
 +
↓
transformation
transformation
 +
strain:XL10-G
 +
 +
</pre>
<html><h3>2010-10-12</h3></html>
<html><h3>2010-10-12</h3></html>
-
Ligation
+
<pre>Ligation
-
 
+
-----------------------
BBa_R0061(100 ng)  1 µL
BBa_R0061(100 ng)  1 µL
BBa_E0240 (100 ng) 3 µL
BBa_E0240 (100 ng) 3 µL
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T4 DNA Ligase      1 µL
T4 DNA Ligase      1 µL
dH<sub>2</sub>O    4 µL
dH<sub>2</sub>O    4 µL
 +
-----------------------
 +
                  10 µL
 +
↓
 +
transformation
 +
strain:XL10-Gold
 +
 +
</pre>
 +
 +
 +
 +
<html><h3>2010-10-21</h3></html>
-
<html><h3>2010-10-13</h3></html>
+
<pre>
 +
Double transformation
 +
Strain:DH10B
 +
Plasmid:Plux inv-GFP & Plac-LuxR
 +
</pre>
 +
<html><h3>2010-10-22</h3></html>
 +
<pre>
 +
liquid culture
 +
Sample:Plux inv-GFP and Plac-LuxR
 +
LB (Carb-Kan,AHL 0 nM and 1000 nM ) 1mL
 +
37℃ 12 h 200 rpm
 +
</pre>
 +
<html><h3>2010-10-26</h3></html>
 +
<pre>
 +
Spin-dawn and observed the pellet
 +
</pre>
　
　

Latest revision as of 03:21, 28 October 2010




 

 





Characterization of lux inverter

2010-10-8

transformation

Plasmid:BBa_R0061
        BBa_E0240
strain:XL10-G


2010-10-9

Mini-prep (Sigma Aldrich)

BBa_R0061
BBa_E0240

digestion
--------------------
BBa_R0061      20 µL
10×Buffer      3 µL
SpeI　　　　　  1 µL
PstI　　　　　  1 µL
NFW             5 µL
--------------------
　　　　　　　 30 µL

--------------------
BBa_E0240      20 µL
10×Buffer      3 µL
XbaI            1 µL
PstI　　　　　  1 µL
NFW             5 µL
--------------------
               30 µL
↓
gel extraction


2010-10-11

Ligation
-----------------------
BBa_R0061 (70 ng)  1 µL       
BBa_E0240 (100 ng) 2 µL
ATP (10 mM)        1 µL
10×Buffer         1 µL
T4 DNA Ligase      1 µL
NFW                4 µL
-----------------------
                  10 µL
↓
transformation
strain:XL10-G

2010-10-12

Ligation
-----------------------
BBa_R0061(100 ng)  1 µL
BBa_E0240 (100 ng) 3 µL
ATP (10 mM)        1 µL
T4 DNA Ligase      1 µL
dH<sub>2</sub>O    4 µL
-----------------------
                  10 µL
↓
transformation
strain:XL10-Gold


2010-10-21

Double transformation

Strain:DH10B
Plasmid:Plux inv-GFP & Plac-LuxR

2010-10-22

liquid culture
Sample:Plux inv-GFP and Plac-LuxR

LB (Carb-Kan,AHL 0 nM and 1000 nM ) 1mL
37℃ 12 h 200 rpm

2010-10-26

Spin-dawn and observed the pellet

　

Retrieved from "http://2010.igem.org/Team:Chiba/Notebook_3"