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.