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This is particularly striking when examining the median methylation value for each CpG site (data not shown).
The median methylation value in the respective patient group was used as the cut point for dichotomization.
From this list of 86 CpGs, we calculated 6 centroids (i.e. profiles consisting of the median methylation value for each of the 86 CpGs) for each of the 6 methylation groups.
The methylation value for each such TSS was taken to be the median methylation value of all of the CpGs within this region with at least 5X coverage, using at least three of the four donors in each state.
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Calculating scores Median methylation values of never smokers taken from previously reported data were used as reference values (supplementary Table 2, as above [ 7]).
Exact binomial tests within these 3 strata (FDR adjusted) were conducted to determine if any of the strata had a significant excess of above or below median methylation values.
Higher median methylation values were seen for all four genes with larger tumor-size (T2, 2 5 cm) than with smaller tumor-size, (T1,< 2 cm) and with aneuploid compared to the diploid cancers (data not shown).
Illumina Methylation Analyzer (IMA) defines for each gene 11 region categories (TSS1500, TSS200, 5′ UTR, 1st EXON, GENE BODY, 3′ UTR, ILSAND, NSHORE, SSHORE, NSHELF, SSHELF) and calculates their mean or median methylation values, which are then compared between the samples under analysis [ 16].
Bedops was used to annotate each CGI from [ 10] with the number of assayed CpGs and their median percent methylation value.
The median relative methylation values found in adN and TU tissues corresponded to 0.087% and 0.634% indicating a 7.3-fold average increase in highly methylated sequences in tumors.
A clear shift towards lower β-values was observed in TAFs compared to CFs (Wilcoxon test, P < 0.001), with a 19.4% reduction of median DNA methylation values in TAFs (dashed vertical lines in Figure 1C).
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