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1. There are 12 different PCR MasterMixes designed to provide a broad range of conditions for a specific DNA template. Thus the preparation of a batch mixture containing DNA template, forward primer, reverse primer, and nuclease-free H2O is recommended to minimize variations caused by pipetting. The PCR set-up for 1 reaction and for 14 reactions (12 reactions for 12 MasterMixes and 2 extra reaction volumes to account for pipetting loss) are as follows: - DNA Template ~100 ng (1 rxn) ~1.4 μg (14 rxn) ~2 ng/μl (Final Conc.) - Forward Primer (10 μM) 1-2.5 μl (1 rxn) 14-35 μl (14 rxn) 200-500 nM (Final Conc.) - Reverse Primer (10 μM) 1-2.5 μl (1 rxn) 14-35 μl (14 rxn) 200-500 nM (Final Conc.) - Nuclease-free H2O Up to 25 μl (1 rxn) Up to 350 μl (14 rxn)2. Mix all components well and aliquot 25 μl to each of the 12 PCR tubes on ice (50 μl reaction volume). Then add 25 μl of each individual PCR-SureTM MasterMix to the 12 PCR tubes.