Difference between revisions of "Team:Paris Bettencourt/Protocols/PCR"

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   <tr>
 
   <tr>
 
     <td>0:30</td>
 
     <td>0:30</td>
−
     <td>64</td>
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     <td>XX°C</td>
 
     <td>annealing</td>
 
     <td>annealing</td>
 
   </tr>
 
   </tr>
 
    
 
    
 
   <tr>
 
   <tr>
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     <td>1:00</td>
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     <td>1 minute/kb</td>
 
     <td>72</td>
 
     <td>72</td>
 
     <td>extension</td>
 
     <td>extension</td>
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     <td>forever</td>
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     <td>infinite hold</td>
 
     <td>12</td>
 
     <td>12</td>
 
     <td>storage</td>
 
     <td>storage</td>

Revision as of 16:03, 1 September 2015

PCR protocol (Phusion)
  • Prepare the following mix
    • 2µL primer Forward (10 µM)
    • 2µL primer Reverse (10 µM)
    • 0.1 to 1 ng of template DNA
    • up to 50µL of DNase/RNase-free water
    • 25µL of Life Technologies Phusion High-Fidelity PCR Master Mix with HF Buffer
  • Launch the PCR using the following mix
    time (min) temperature (°C) function
    3:00 98 melting
    0:30 98 melting
    0:30 XX°C annealing
    1 minute/kb 72 extension
    10:00 72 extension
    infinite hold 12 storage