Older PCR protocol

You should first run qPCR to adjust template concentration (see real-time PCR).

4X linear PCR:

ReagentInitialFinal1X reaction (50 μl)4.5X (μl)
Kapa Buffer A10x1x522.5
Forward10 µM0.4 µM29
Reverse (T7)10 µM0.4 µM29
dNTP mix10 mM0.4 mM29
Raw library2 ng/µL0.2 ng/µL522.5
Water 33.5150.75
Kapa Taq5 U/µL0.5 U/µL0.52.25
4X linear PCR

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