Fig. 4

Technical sensitivity and coverage. Sensitivity of the MBPS assay was assessed using methylated-control DNA at 0%, 1% and 5% expected levels of methylation, across three separate sequencing runs. Two representative amplicons (amplicons 44 and 55) are shown. Observed methylation (%) (y-axis) is plotted against expected methylation (%) (x-axis). a Both representative amplicons show statistically significant detection of both 1% and 5% methylation. b, c Down-sampling sequencing coverage at 100x, 1000x, 10,000x and 100,000x shows that the sensitivity of detection of methylation improves with increased coverage. 1000x coverage and greater enables detection of 1% methylation that is statistically significant. ns = not significant (p ≥ 0.05), * = 0.01 ≤ p < 0.05, ** = 0.001 ≤ p < 0.01, *** = 0.0001 ≤ p < 0.001