Team:SCU-China/Linkage

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             <li><a href="https://2014.igem.org/Team:SCU-China/Attributions">Attributions</a></li>
             <li><a href="https://2014.igem.org/Team:SCU-China/Attributions">Attributions</a></li>
             <li><a href="https://2014.igem.org/Team:SCU-China/Team">Team</a></li>
             <li><a href="https://2014.igem.org/Team:SCU-China/Team">Team</a></li>
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             <li class="dropdown   active"><a href="#" class="dropdown-toggle" data-toggle="dropdown">Notbook<span class="caret"></span></a>
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             <li class="dropdown active"><a href="#" class="dropdown-toggle" data-toggle="dropdown">Notebook<span class="caret"></span></a>
               <ul class="dropdown-menu" role="menu">
               <ul class="dropdown-menu" role="menu">
                 <li class="dropdown-header">Notebook</li>
                 <li class="dropdown-header">Notebook</li>
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                <li><a href="https://2014.igem.org/Team:SCU-China/Transmitter">Notebook of Transmitter</a></li>
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                <li><a href="https://2014.igem.org/Team:SCU-China/Biobricks">Notebook of Biobricks</a></li>
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<li><a href="https://2014.igem.org/Team:SCU-China/Transmitter">Notebook of Transmitter</a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/Effector">Notebook of Effector</a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/Effector">Notebook of Effector</a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/Sensor">Notebook of Sensor</a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/Sensor">Notebook of Sensor</a></li>
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                 <li class="dropdown-header">Method</li>
                 <li class="dropdown-header">Method</li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/Prep">Bacterial Genomic DNA Prep</a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/Prep">Bacterial Genomic DNA Prep</a></li>
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                 <li><a href="https://2014.igem.org/Team:SCU-China/Digestion">Digestionk</a></li>
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                 <li><a href="https://2014.igem.org/Team:SCU-China/Digestion">Digestion</a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/GelExtraction">Gel Extraction </a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/GelExtraction">Gel Extraction </a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/Linkage">Linkage</a></li>
                 <li><a href="https://2014.igem.org/Team:SCU-China/Linkage">Linkage</a></li>

Latest revision as of 18:55, 17 October 2014

Linkage Protocol

According to the advice of IGEM Competition, there are two linkage methods that we use in experiment. The first one is “3A” Assembly and the second is “Traditional” Assembly.

For “3A” Assembly, we set up the following system (20 μl):

Linear Backbones from IGEM Competition( with K+, C+, A+, or T+)

About 100 ng

Insert DNA 1

The molar ratio of Insert DNA to Backbone is 3:1 to 5:1

Insert DNA 2

Same to Insert DNA 1

10 x T4 DNA Ligase Buffer

2 μl

T4 DNA Ligase

1 μl

H2O

Up to 20 μl

For “Traditional” Assembly, we set up the following system (20 μl):

Linear Backbones from IGEM Competition( with K+, C+, A+, or T+)

About 100 ng

Insert DNA

The molar ratio of Insert DNA to Backbone is 3:1 to 5:1

10 x T4 DNA Ligase Buffer

2 μl

T4 DNA Ligase

1 μl

H2O

Up to 20 μl

Sichuan university