Difference between revisions of "Team:UiOslo Norway/Experiments/PCR taq"

 
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Using the guidelines of the provided Taq DNA Polymerase with Standard Taq Buffer (M0273)
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<a href="https://www.neb.com/protocols/1/01/01/taq-dna-polymerase-with-standard-taq-buffer-m0273" >
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protocol.
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   <li><p><b>Reaction Setup:</b></br>
 
   <li><p><b>Reaction Setup:</b></br>
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     <td>95 °C</td>
 
     <td>95 °C</td>
 
     <td>30 seconds /5 Minutes for Colony PCR
 
     <td>30 seconds /5 Minutes for Colony PCR
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   </tr>
 
   <tr>
 
   <tr>
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     <td>35 Cycles</td>
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     <td rowspan="3"></br></br>35 Cycles</td>
 
     <td>95 °C</td>
 
     <td>95 °C</td>
 
     <td>30 seconds</td>
 
     <td>30 seconds</td>
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    </tr>
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    <td>45 °C – 72 °C</td>
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    <td>60 seconds</td>
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    <td>68 °C</td>
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    <td>1 minute / 1 kb</td>
 
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</div></div> <!--These are the closing tags for div id="mainContainer" and div id="contentContainer". The corresponding opening tags appear in the template that is {{included}} at the top of this page.-->
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<div class="clear"></div>
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<!---------------sponsors------------------------->
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<center><h1>iGEM UiOslo 2015 is sponsored by:</h1>
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<hr style="height:2px;border:none;color:black;background-color:black;" />
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<img src="https://static.igem.org/mediawiki/2015/a/ac/UiOslo_Statoil.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/e/e0/UiOslo_uio_logo.png" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/0/08/UiOslo_Enova1.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/e/eb/UiOslo_GATC-Biotech.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/4/4a/UiOslo_IDT.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/d/d9/UiOslo_Norwegian_Biochemical_Society.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/f/f0/UiOslo_SnapGene.jpg" Height="60px" hspace="15">
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</center>
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<hr style="height:2px;border:none;color:black;background-color:black;" />
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<!--------end sponsors---------------------------->
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</body>
 
</html>
 
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Latest revision as of 12:10, 18 September 2015

PCR with Taq DNA Polymerase (NEB):

Back to Protocols


Using the guidelines of the provided Taq DNA Polymerase with Standard Taq Buffer (M0273) protocol.

  1. Reaction Setup:

    2.5 µl 10X Standard Taq Reaction buffer
    2.5 µl dNTPS [2 mM]
    0.5 µl Forward Primer [10 µM]
    0.5 µl Reverse Primer [10 µM]
    1 µl Template DNA
    0.125 µl Taq DNA Polymerase
    17.875 µl MilliQ Water

    25 µl Total Reaction


  2. PCR Program:

    Step Temperature Time
    Initial Denaturation 95 °C 30 seconds /5 Minutes for Colony PCR


    35 Cycles    		 95 °C 30 seconds
    45 °C – 72 °C 60 seconds
    68 °C 1 minute / 1 kb
    Final Extension 68 °C 5 minutes
    Hold 4 °C forever

    3. iGEM UiOslo 2015 is sponsored by: