Table 1.

Preparation of Multiplex STEP 1 Reaction Mix for One Sample, 12 genes

Multiplex STEP 1 reaction mix	1×
	Master mix (μL)	Final concentration
10× MSP	5.0	1×
25 mM dNTP	2.5	1.25 mM
Platinum Taq (5 U/μL)	2.0	0.2 U/μL
Water	6.5
Subtotal μL	16.0
For each of 12 genes:
External primer 100 ng/μl forward	1.0 × 12 genes	2 ng/μL per gene
External primer 100 ng/μl reverse	1.0 × 12 genes	2 ng/μL per gene
Subtotal μL	24.0
DNA (bisulfite-treated) ≤ 40 ng total	10.0
Total μL	50