Team:USTC CHINA/Notebook/Protocols/10*S base

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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">notebook</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">protocols</a></div></div>
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<div id="breadcrumb"><a href="https://2013.igem.org/Team:USTC_CHINA">Home</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook">notebook</a> &gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols">protocols</a>&gt; <a href="https://2013.igem.org/Team:USTC_CHINA/Notebook/Protocols/10*S base">protocols 10*S base</a></div></div>
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<h1>Gel Extraction</h1>
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<h1>10*S base</h1>
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<p>Performed with AxyPrep 96-well DNA Gel Extraction Kit (type: AP-96-GX)
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<p><table width="580" border="2">
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Protocol
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  <tr>
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1. Excise gel slice containing DNA fragment of interest.
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    <td>Ammonium Sulfate</td>
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2. Add 3×sample volume of Buffer DE-A.
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    <td>2g</td>
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Incubate at 75° C for 15-20 min or until gel melts completely.
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  </tr>
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Add 0.5 × Buffer DE-A volume of Buffer DE-B.
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  <tr>
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<img src="https://static.igem.org/mediawiki/2013/4/4d/2013igemustc-china_Protocol1.png" width="580" height="200" />  
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    <td>Dipotassium Phosphate</td>
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3. Binding sample DNA(Centrifuge at 5,000 - 6,000×g for 5 min)
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    <td>14g</td>
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  </tr>
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4. Add 800 μl of Buffer W2 (5,000 - 6,000×g for 1 min)
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  <tr>
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Repeat wash with Buffer W2
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    <td>Potassium Dihydrogen Phosphate</td>
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Centrifuge empty plate for 5 min at 5,000 - 6,000×g to remove residue W2.
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    <td>6g</td>
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  </tr>
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5. Elute with 50 μl of Eluent or Deionized Water.(5,000 - 6,000×g, 5min)
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  <tr>
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    <td>Sodium Citrate</td>
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    <td>1g</td>
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  </tr>
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</table>
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Add deionized water and fix the volume at 100ml, and add 0.1mol/L (NH4)2SO4 after autoclaving.</br>
   
   

Revision as of 15:22, 25 September 2013

10*S base

Ammonium Sulfate 2g
Dipotassium Phosphate 14g
Potassium Dihydrogen Phosphate 6g
Sodium Citrate 1g
Add deionized water and fix the volume at 100ml, and add 0.1mol/L (NH4)2SO4 after autoclaving.

notebook

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