Table 5. Library construction PCR conditions.

Reaction mixture
		Volume
2× PCR mix (KAPA HiFi HotStart ReadyMix)		20 μl
P5 end primer (10 μM)		1 μl
P7 end primer (10 μM)		1 μl
Column-purified NASBA product (~3.5 ng dsDNA)		4 μl
Nuclease-free water		14 μl
Thermocycling conditions
Temperature	Time	Cycle number
95°C	3 min	1
98°C	20 s	15
60°C	15 s
72°C	30 s
72°C	4 min	1