Analysis on the Susceptibility Genes in Two Chinese Pedigrees with Familial Parkinson's Disease

Table 1

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