Team:Groningen/protocols/Ligation

From 2013.igem.org

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<h1>Ligation</h1>
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<h3>Materials:</h3>
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<ul type="square">
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<li>H<sub>2</sub>O</li>
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<li>1.5ml tubes</li>
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<li>Ligation buffer</li>
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<li>Ligase</li>
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<li>Insert</li>
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<li>Vector</li>
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</ul>
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<h3>Reaction:</h3>
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<table border = "1">
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<tr><td>Component</td>
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<td>30&#956;l reaction</td>
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<td>Final concentration</td>
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</tr>
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<tr><td>H<sub>2</sub>O</td>
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<td>up to 30&#956;l</td>
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<td></td>
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</tr>
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<tr><td>Buffer 10x</td>
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<td>3&#956;l</td>
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<td>1x</td>
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</tr>
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<tr><td>DNA</td>
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<td>4&#956;l</td>
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<td>up to 1&#956;g</td>
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</tr>
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<tr><td>Enzyme/s</td>
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<td>0.5&#956;l</td>
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<td></td>
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</tr>
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</table>
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<br>All the reagents are added following the order listed in the table above.
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<br>After the reaction is ready mix the content of the tube and spin it down.
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<br>The tubes are incubated for 1h at 37&deg;C.
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<br>
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<br>Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf
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Revision as of 09:56, 26 July 2013

Ligation

Materials:

  • H2O
  • 1.5ml tubes
  • Ligation buffer
  • Ligase
  • Insert
  • Vector

Reaction:

Component 30μl reaction Final concentration
H2O up to 30μl
Buffer 10x 3μl 1x
DNA 4μl up to 1μg
Enzyme/s 0.5μl

All the reagents are added following the order listed in the table above.
After the reaction is ready mix the content of the tube and spin it down.
The tubes are incubated for 1h at 37°C.

Reference: http://www.thermoscientificbio.com/uploadedFiles/Resources/fast-digestion-dna.pdf