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<- map_dfr( # plot + annotations p <- ggplot(dfg, aes(x = fract, y = n_sites, group = minDepth, color = minDepth)) + - geom_line() + # highlighting the minDepth threshold of choice geom_line(data = dfg[minDepth == 5, ], aes(x = fract, y = n_sites), color = "red", size = 1.5) + + geom_line() + scale_color_viridis_c() + coord_cartesian(ylim = c(0, 2E6)) + # highlighting the fract threshold used @@ -74,3 +74,61 @@ p <- ggplot(dfg, aes(x = fract, y = n_sites, group = minDepth, color = minDepth) ggsave("minimum_coverage_thresholding.png", p, width = 7, height = 5) +# histograms ----------------------- +#before filtering +dfph <- study_whole +dfph <- dfph[Cyt != 0 | Thy != 0] +dfph[, count := Cyt + Thy] +sampleOrder <- dfph[, mean(count), by = "label"][order(V1), ]$label +dfph <- dfph[, .N, by = c("label", "count")] +dfph <- dfph[order(label, count), ] +dfph$step <- "1- Before filtering" + +#after removing minDepth +pos_to_remove_minDepth <- BSFilterMinDepth(study_DT = study_whole, minDepth = 5, sampleFraction = 0.8) +study_whole <- study_whole[!pos_to_remove_minDepth] +dfph2 <- study_whole +dfph2 <- dfph2[Cyt != 0 | Thy != 0] +dfph2[, count := Cyt + Thy] +dfph2 <- dfph2[, .N, by = c("label", "count")] +dfph2 <- dfph2[order(label, count), ] +dfph2$step <- "2- After MinDepth" +dfph <- rbind(dfph, dfph2) + +#after removing maxDepth +pos_to_remove_maxDepth <- BSFilterMaxDepth(study_DT = study_whole, maxDepth = 100) +study_whole <- study_whole[!pos_to_remove_maxDepth] +dfph2 <- study_whole +dfph2 <- dfph2[Cyt != 0 | Thy != 0] +dfph2[, count := Cyt + Thy] +dfph2 <- dfph2[, .N, by = c("label", "count")] +dfph2 <- dfph2[order(label, count), ] +dfph2$step <- "3- After MaxDepth" +dfph <- rbind(dfph, dfph2) + +#normalisation +study_whole <- BSCoverageNorm(study_DT = study_whole) +dfph2 <- study_whole +dfph2 <- dfph2[Cyt != 0 | Thy != 0] +dfph2[, count := Cyt + Thy] +dfph2 <- dfph2[, .N, by = c("label", "count")] +dfph2 <- dfph2[order(label, count), ] +dfph2$step <- "4- After Normalisation" +dfph <- rbind(dfph, dfph2) + +#mutli-facet plot +p <- ggplot(dfph, aes(x = factor(label, levels = sampleOrder), y = N, fill = count)) + + geom_col() + + scale_fill_viridis_c(limits = c(1, 100), oob = scales::squish) + + labs( + title = "CpG filtering", + x = "Sample", + y = "Number of CpG sites", + fill = "Coverage" + ) + + scale_y_continuous(labels = scales::comma) + + theme_bw(base_size = 14) + + theme(axis.text.x = element_text(angle = 90, hjust = 1, size = 8)) + + facet_wrap(~step, ncol = 4) + +ggsave("all_thresholding_steps.png", p, width = 12, height = 4) -- GitLab