Table 1.

1. PCR
DW	15.1 μ L	14.1 μ L
DMSO	—	1.0 μ L	94°C 10 min–94°C 30 sec↓–72°C 10 min–4°C
10 × PCR Buf	2.0 μ L	2.0 μ L	52°C to 60°C 30 sec↓
2.5 mM dNTP	1.0 μ L	1.0 μ L	72°C 45 sec↓
3.5 U/ul HiFi	0.3 μ L	0.3 μ L	×40 cycles
10 pm For primer	0.5 μ L	0.5 μ L
10 pm Rev primer	0.5 μ L	0.5 μ L
DNA	0.6 μ L	0.6 μ L
Total	20.0 μ L	20.0 μ L
2. Sequence
DW	6.3 μ L
1 × Seq Buf	1.0 μ L		96°C 1 min–96°C 10 sec↓–4°C
BigDye	0.2 μ L		60°C 4 min↓
0.8 pm For primer (or Rev primer)	2.0 μ L		×30 cycles
Template DNA	0.5 μ L
Total	10.0 μ L
3. Denature			95°C 5 min–4°C
4. QPCR
THUNDERBIRD SYBR qPCR Mix	9 μ L		95°C 1 min–95°C 15 sec
Forward Primer	6 pmol		57°C 20 sec
Reverse Primer	6 pmol		72°C 40 sec
50× ROX reference dye	0.36 μ L		×40 cycles
Template DNA	1 μ L
	Upto18 microlitter