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Table 2 PCR conditions and cycling protocol with PerfectTaq DNA polymerase™ (5 Prime)

From: Where Do I Come From? Using Student’s Mitochondrial DNA to Teach About Phylogeny, Molecular Clocks, and Population Genetics

Components Total volume (μl) Final concentration
10× PCR Buffer 5
dNTP mix (ten mM each) 1 200 μM
Primers (ten μM) each, Table 1 1 each 0.2 μM each
Water (RNase free) 31.75
PerfectTaq enzyme 0.25 1.25 units
DNA 10 ~20–200 ηg
Total volume 50
  1. Initial denaturation of three min at 94 °C followed by 35 cycles of 30 s at 94 °C, 30 s at 51 °C, and 60 s at 72 °C and a final extension of ten min at 72 °C