Team:Alberta/Project/AnchTermAnneal

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     <h1>Annealing Anchor and Terminator Bytes</h1>
     <h1>Annealing Anchor and Terminator Bytes</h1>
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Instructions on how to anneal the complementary strands to obtain the same concentrations we used in our experiments (10uM).
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Instructions on how to anneal the complementary strands to obtain the same concentrations we used in our experiments (10μM).
<h2>What you will need:</h2>
<h2>What you will need:</h2>
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<h2>Procedure:</h2>
<h2>Procedure:</h2>
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To make a 10 μM Anchor solution with an A(B) overhang one must anneal Anchor_A(Anchor_B) and Anchor_Comp. To make a 10 μM Terminator solution with an A(B) overhang one must anneal Term_A(Term_B) and Term_Comp.
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<ul>
<ul>
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    <li>Pipet 150 uL 50% glycerol into 3* properly labeled 1.5 ml screw cap tubes.
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<li>Combine 10 μL of Anchor/Term_(A or B) and Anchor/Term_Comp 100 μM stock solutions into an eppendorf tube.
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    <li>Add 350 uL of overnight culture to each tube.
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<li>Add 80 μL water or TE, mix.  
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    <li>Pipet up and down to gently mix.
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<li>Heat solution to 95C for 2 minutes.
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    <li>Flash freeze in liquid N2 or dry ice/methanol bath
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<li>Let cool to room temperature slowly.
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    <li>Place in -80 C freezer when frozen.  
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</ul>
</ul>

Latest revision as of 16:52, 21 October 2009

University of Alberta - BioBytes










































































































Annealing Anchor and Terminator Bytes

Instructions on how to anneal the complementary strands to obtain the same concentrations we used in our experiments (10μM).

What you will need:

  • Anchor oligos (complementary)
  • Terminator oligos (complementary)

Procedure:

To make a 10 μM Anchor solution with an A(B) overhang one must anneal Anchor_A(Anchor_B) and Anchor_Comp. To make a 10 μM Terminator solution with an A(B) overhang one must anneal Term_A(Term_B) and Term_Comp.
  • Combine 10 μL of Anchor/Term_(A or B) and Anchor/Term_Comp 100 μM stock solutions into an eppendorf tube.
  • Add 80 μL water or TE, mix.
  • Heat solution to 95C for 2 minutes.
  • Let cool to room temperature slowly.

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