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. Author manuscript; available in PMC: 2019 Jul 2.
Published in final edited form as: Methods Mol Biol. 2018;1708:645–663. doi: 10.1007/978-1-4939-7481-8_33
Initial denaturation 95 °C 2 min
Denaturation 95 °C 30 s
Annealing Primer dependent (see Note 28) 30 s
Extension 72 °C 1 min
40 cycles
Final extension 72 °C 7 min
Hold 4°C