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.