Team:Chiba/Notebook 3

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<li><a href="https://2010.igem.org/Team:Chiba"><span>Home</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 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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<li><a href="https://2010.igem.org/Team:Chiba/Notebook"><span>Notebook</span></a>
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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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                                <!-- CSS Tabs -->
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<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>
 +
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-
'''Lux inverter'''
 
-
== Headline text ==
 
-
BBa_R0061&BBa_E0240
+
<!--- 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>
 +
 
 +
<html><h3>2010-10-9</h3></html>
 +
 
 +
<pre>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
 +
 
 +
</pre>
 +
 
 +
 
 +
<html><h3>2010-10-11</h3></html>
 +
<pre>
 +
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
 +
 
 +
</pre>
 +
 
 +
<html><h3>2010-10-12</h3></html>
 +
 
 +
<pre>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
 +
 
 +
</pre>
 +
 
 +
 
 +
 
 +
<html><h3>2010-10-21</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"