Team:Tuebingen/Notebook/Protocols/ligation

From 2013.igem.org

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<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 16px">Back to Protocols</a>
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<a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a>
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<p>&nbsp;</p>
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<h3>Reagents</h3>
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    <td style="text-align: center">20 - 50 ng</td>
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    <td>Linearized vector</td>
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  </tr>
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    <td style="text-align: center">1 µL</td>
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    <td>10x Ligase buffer</td>
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    <td style="text-align: center">1 µL</td>
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    <td>T4 Ligase</td>
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  </tr>
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  <tr>
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    <td style="text-align: center">5 µl (up to 5:1 molar ratio insert to vector) </td>
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    <td>Insert DNA</td>
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    <td style="text-align: center">to 10 µL</td>
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    <td>Aqua dest.</td>
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<p>&nbsp;</p>
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<h3>Procedure</h3>
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<p>Dissolve all reagents in 1000 mL Aqua dest., adjust pH to 7.0 (with NaOH). Sterilize by autoclaving. After autoclaving add 5 mL MgCL2 (c = 2 M).</p>
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Revision as of 00:17, 4 October 2013

Return to iGEM Main Page.

Ligation
Back to Protocols

 

Reagents

20 - 50 ng Linearized vector
1 µL 10x Ligase buffer
1 µL T4 Ligase
5 µl (up to 5:1 molar ratio insert to vector) Insert DNA
to 10 µL Aqua dest.

 

Procedure

Dissolve all reagents in 1000 mL Aqua dest., adjust pH to 7.0 (with NaOH). Sterilize by autoclaving. After autoclaving add 5 mL MgCL2 (c = 2 M).


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