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

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   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
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     <td rowspan="2">35 Cycles</td>
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     <td rowspan="3">35 Cycles</td>
 
     <td>95 °C</td>
 
     <td>95 °C</td>
 
     <td>30 seconds</td>
 
     <td>30 seconds</td>
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     <td>45 °C – 72 °C</td>
 
     <td>45 °C – 72 °C</td>
 
     <td>60 seconds</td>
 
     <td>60 seconds</td>
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  </tr>
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    </tr>
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    <tr>
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    <td>68 °C</td>
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    <td>1 minute / 1 kb</td>
 
   </tr>
 
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Revision as of 10:01, 14 August 2015

PCR with Taq DNA Polymerase (NEB):

Back to Protocols


  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