Table 3.

MethyLight qPCR and dPCR

Run no.	MID no.	% methylation
		Expected (× replicate no.)	Observed
1	1	100	100
	2	90	96.4
	3	75	87.7
	9	50 × 1	85.3
	4	50 × 2	60.2
	7	50 × 3	64.3
	5	25 × 1	35.6
	10	25 × 2	38.7
	8	10	24.9
	6	0	0.2
2	7	100	99.7
	3	90	94.8
	8	75	91.8
	9	50 × 1	84.5
	10	50 × 2	61.5
	6	50 × 3	74.5
	1	25 × 1	29.8
	4	25 × 2	28.2
	5	10	15.7
	2	0	0.6

The % methylation determined by bisulfite amplicon NGS using Multiplex Identifier (MID) fusion primers to amplify samples from a panel of methylated/unmethylated DNA standards mixed in a range of known ratios. The 25% and 50% methylated samples were replicated (× 1-3) within each of two runs.