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

 
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Line 12: Line 12:
 
         <li>vector 100ng
 
         <li>vector 100ng
 
         <li>insert 300ng
 
         <li>insert 300ng
−
         <li>T4 DNA ligase buffer 10X
+
         <li>2µL T4 DNA ligase buffer 10X
−
         <li>T4 DNA ligase
+
         <li>1µL T4 DNA ligase
 
         <li>up to 20µL of water
 
         <li>up to 20µL of water
 
       </ul>
 
       </ul>
−
       <li>incubate 30 to 40 minutes at room temperature
+
       <li>incubate 10 to 15 minutes at 22°C
 +
      <li>incubate 5 minutes at 70°C
 +
 
 
     </ul>
 
     </ul>
 
     </td>
 
     </td>
 
   </tr>
 
   </tr>
 
</table>
 
</table>

Latest revision as of 18:36, 7 September 2015

Ligation Protocol
  • Mix the following
    • vector 100ng
    • insert 300ng
    • 2µL T4 DNA ligase buffer 10X
    • 1µL T4 DNA ligase
    • up to 20µL of water
  • incubate 10 to 15 minutes at 22°C
  • incubate 5 minutes at 70°C