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  • End-point PCR Using EpiMark® Bisulfite Conversion Kit (E3318)

    Protocol

    1. Add the following components to a 0.2 ml PCR reaction tube:
      PCR Component 25 μl Reaction 50 μl Reaction Final
      Concentration
      5X EpiMark™
      Hot Start Taq Reaction
      Buffer
      5μl 10μl 1X
      10 mM dNTPs 0.5 μl 1 μl 200 µM
      10 μM Forward Primer 0.5 μl 1 μl 0.2 µM
      10 μM Reverse Primer 0.5 μl 1 μl 0.2 µM
      Template DNA 1-6 μl 1-6 μl up to 1 μg
      EpiMark™ Hot Start Taq
      DNA Polymerase
      0.125 μl 0.25 μl 1.25 units/50 μl
      PCR reaction
      Nuclease-free water up to 25 μl up to 50 μl  


    2. Gently mix the reaction. If necessary, collect all liquid to the bottom of the tube by a quick spin. Overlay the sample with mineral oil if using a thermocycler without a heated lid.

    3. Transfer PCR tubes from ice to a thermocycler with the block preheated to 94°C and start the cycling program.