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

 
Line 16: Line 16:
 
           <li>complete to 50µL of DNase/RNase-free water</li>
 
           <li>complete to 50µL of DNase/RNase-free water</li>
 
         </ul>
 
         </ul>
−
       <li>Launch the PCR using the following mix
+
       <li>Launch 30 to 35 PCR cycles using the following parameters:<br>
  
 
           <table style="width:25%">
 
           <table style="width:25%">

Latest revision as of 16:05, 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
    • 25µL of Life Technologies Phusion High-Fidelity PCR Master Mix with HF Buffer
    • complete to 50µL of DNase/RNase-free water
  • Launch 30 to 35 PCR cycles using the following parameters:
    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