Team:Tuebingen/Notebook/Protocols/pgem-ligation

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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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<h3>Reagents</h3>
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    <td style="text-align: center">5 µL</td>
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    <td>2x Rapid Ligation Buffer</td>
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    <td style="text-align: center">0.5 µL (25 ng)</td>
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    <td>pGEM vector</td>
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    <td style="text-align: center">3.5 µL</td>
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    <td>PCR</td>
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    <td style="text-align: center">10 mL</td>
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    <td>KCl (c = 250 mM)</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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Latest revision as of 12:09, 4 October 2013

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pGEM Ligation

Reagents

5 µL 2x Rapid Ligation Buffer
0.5 µL (25 ng) pGEM vector
3.5 µL PCR
10 mL KCl (c = 250 mM)

 

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).

Back to Protocols