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Table 2 Methylight qPCR and dPCR

From: Quantification of epigenetic biomarkers: an evaluation of established and emerging methods for DNA methylation analysis

Expected methylation (%) Average methylation ± standard deviation (%)
qPCR dPCR
P14_M P14_M P14_M P14_M2 P14_M + P14_M2
Singleplex Singleplex Duplex Duplex Duplex
100 100* 100 100 100 100
90 78 ± 4¥ 77 ± 27 99 ± 19* 86 ± 20 92 ± 19
75 66 ± 18* 81 ± 13 86 ± 20 77 ± 17 81 ± 19
50 49 ± 10¥,+ 52 ± 16 58 ± 10*,¥ 57 ± 28* 56 ± 18*
25 28 ± 4 21 ± 7 31 ± 5 28 ± 2 29 ± 4
10 13 ± 3+ 17 ± 4 18 ± 6¥ 9 ± 15* 13 ± 10*
0 0 0 0 0 0
  1. The average % methylation calculated from three independent qPCR and dPCR measurements of three independent bisulfite conversions of a panel of methylated/unmethylated DNA standards; Data for dPCR show measurements from the P14_M assay used in singleplex and for the P14_M and P14_M2 assays used together in duplex, showing the % methylation for each assay when analysed individually and with the estimated targets combined (P14_M + P14_M2). Symbols denote statistical comparisons using a One-Way ANOVA test. All comparisons between samples with expected differences in methylation of ≥ 50% were significant at the level of p < 0.05 (with the exception of 50% vs. 100% methylation with MethyLight P14_M2 Duplex) and are not shown; all comparisons between samples of differences between 0-40% expected methylation that are significant (p < 0.05) are shown with pairs of the same symbols (*,¥,+,‡) denoting which two samples were compared, e.g. for dPCR P14_M2 duplex, only the 10% and 50% methylated (expected % methylation) data points were significantly different from each other at the level of p < 0.05.