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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: Methods Mol Biol. 2014;1086:53–77. doi: 10.1007/978-1-62703-667-2_4

Table 2.

PCR program used to assemble the DNA sequence that will be transcribed into the desired RNA. Annealing temperatures may have to be adjusted depending on the melting temperatures of each primer in the assembly

Steps Time/cycles Temperature
Denaturation 30 s 98 °C
Denaturation 10 s 98 °C
Annealing 30 s 64 °C
Extension 30 s 72 °C
Repeat steps 2–4 29 cycles more
Polymerization 10 min 72 °C