Difference between revisions of "Team:Stony Brook/Protocols"

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<h2> Lab Protocols</h2>
 
<h2> Lab Protocols</h2>
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</div>
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<div class="column half_size">
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<h3> Nanodrop </h3>
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<ol>
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<li> Open "Nucleic Acids" </li>
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<li> Drop water on platform. Close it and blank. </li>
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<li> Blank system with the solvent used for protocol </li>
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<li> Drop 1ul of sample on platform, close, click "measure"</li>
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<li> Should have two humps </li>
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</ol>
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</div>
  
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<div class="column half_size">
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<h3> Ligation </h3>
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<ol>
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<li><pre>
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10X DNA Ligase Buffer → 2ul
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Vector DNA (4kb) → 50ng
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Insert DNA (1kb) → 37.5ng
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Nuclease-Free Water → fill to 20ul
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T4 DNA ligase → 1ul
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</pre></li>
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<li> Mix by pipetting up and down</li>
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<li> Incubate at 16°C overnight or room temp for 10 minutes </li>
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<li> Heat inactive (heat block) at 65°C for 10 minutes </li>
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<li> Chill on ice and transform 1-5ul of the reaction into 50ul competent cells</li>
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<li> Insert in order of increasing expensiveness </li>
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</ol>
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</div>
  
  

Revision as of 21:09, 14 July 2016

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Lab Protocols

Nanodrop

  1. Open "Nucleic Acids"
  2. Drop water on platform. Close it and blank.
  3. Blank system with the solvent used for protocol
  4. Drop 1ul of sample on platform, close, click "measure"
  5. Should have two humps

Ligation 

  1.  
10X DNA Ligase Buffer → 2ul
Vector DNA (4kb) → 50ng
Insert DNA (1kb) → 37.5ng
Nuclease-Free Water → fill to 20ul
T4 DNA ligase → 1ul
  2. Mix by pipetting up and down
  3. Incubate at 16°C overnight or room temp for 10 minutes
  4. Heat inactive (heat block) at 65°C for 10 minutes
  5. Chill on ice and transform 1-5ul of the reaction into 50ul competent cells
  6. Insert in order of increasing expensiveness