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

Line 38: Line 38:
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td>Jill</td>
+
     <td>Initial Denaturation</td>
−
     <td>Smith</td>
+
     <td>95 °C</td>
−
     <td>50</td>
+
     <td>30 seconds /5 Minutes for Colony PCR
 +
</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td>Eve</td>
+
     <td>35 Cycles</td>
−
     <td>Jackson</td>
+
     <td>95 °C</td>
−
     <td>94</td>
+
     <td>30 seconds</td>
 
   </tr>
 
   </tr>
 
   <tr>
 
   <tr>
−
     <td>John</td>
+
     <td>Final Extension</td>
−
     <td>Doe</td>
+
     <td>68 °C</td>
−
     <td>80</td>
+
     <td>5 minutes</td>
 +
  </tr>
 +
  <tr>
 +
    <td>Hold</td>
 +
    <td>4 °C</td>
 +
    <td>forever</td>
 
   </tr>
 
   </tr>
 
</table>
 
</table>

Revision as of 09:06, 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
    Final Extension 68 °C 5 minutes
    Hold 4 °C forever