annotate scater-plot-dist-scatter.R @ 2:b834074a9aff draft

"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
author iuc
date Thu, 09 Sep 2021 12:23:11 +0000
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1 #!/usr/bin/env Rscript
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3 # Plot the distribution of read counts and feature counts, side by side, then a scatter plot of read counts vs feature counts below
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5 # Load optparse we need to check inputs
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7 library(optparse)
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8 library(workflowscriptscommon)
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9 library(LoomExperiment)
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10 library(scater)
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11 library(ggpubr)
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12 library(scales)
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14 # parse options
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15
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16 option_list <- list(
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17 make_option(
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18 c("-i", "--input-loom"),
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19 action = "store",
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20 default = NA,
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21 type = "character",
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22 help = "A SingleCellExperiment object file in Loom format."
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23 ),
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24 make_option(
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25 c("-o", "--output-plot-file"),
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26 action = "store",
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27 default = NA,
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28 type = "character",
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29 help = "Path of the PDF output file to save plot to."
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30 ),
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31 make_option(
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32 c("-l", "--log-scale"),
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33 action = "store_true",
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34 default = FALSE,
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35 type = "logical",
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36 help = "Plot on log scale (recommended for large datasets)."
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37 )
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38 )
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39
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40 opt <- wsc_parse_args(option_list, mandatory = c("input_loom", "output_plot_file"))
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42 # Check parameter values
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43
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44 if (! file.exists(opt$input_loom)) {
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45 stop((paste("File", opt$input_loom, "does not exist")))
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46 }
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48 # Filter out unexpressed features
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49
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50 sce <- import(opt$input_loom, format = "loom", type = "SingleCellLoomExperiment")
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52 # Do the scatter plot of reads vs genes
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53 total_counts <- sce$total
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54 total_features <- sce$detected
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55 count_feats <- cbind(total_counts, total_features)
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56 cf_dm <- as.data.frame(count_feats)
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57
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58 # Calculate binwidths for reads and features plots. Use 20 bins
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59 read_bins <- max(total_counts / 1e6) / 20
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60 feat_bins <- max(total_features) / 20
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61
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62 plot1 <- qplot(total_counts / 1e6, geom = "histogram", binwidth = read_bins, ylab = "Number of cells", xlab = "Read counts (millions)", fill = I("darkseagreen3")) + ggtitle("Read counts per cell")
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63 plot2 <- qplot(total_features, geom = "histogram", binwidth = feat_bins, ylab = "Number of cells", xlab = "Feature counts", fill = I("darkseagreen3")) + ggtitle("Feature counts per cell")
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64 plot3 <- ggplot(cf_dm, aes(x = total_counts / 1e6, y = total_features)) + geom_point(shape = 1) + geom_smooth() + xlab("Read count (millions)") +
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65 ylab("Feature count") + ggtitle("Scatterplot of reads vs features")
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66 plot4 <- plotColData(sce, y = "subsets_Mito_percent", x = "detected") + ggtitle("% MT genes") + geom_point(shape = 1) + theme(text = element_text(size = 15)) + theme(plot.title = element_text(size = 15)) + xlab("Total features") + ylab("% MT")
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67
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68 if (! opt$log_scale) {
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69 final_plot <- ggarrange(plot1, plot2, plot3, plot4, ncol = 2, nrow = 2)
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70 ggsave(opt$output_plot_file, final_plot, device = "pdf")
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71 } else {
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72 plot1_log <- plot1 + scale_x_continuous(trans = "log10") + scale_y_continuous(trans = "log10")
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73 plot2_log <- plot2 + scale_y_continuous(trans = "log10")
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74 plot3_log <- plot3 + scale_y_continuous(trans = "log10")
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75 plot4_log <- plot4 + scale_y_log10(labels = number)
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76 final_plot_log <- ggarrange(plot1_log, plot2_log, plot3_log, plot4_log, ncol = 2, nrow = 2)
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77 ggsave(opt$output_plot_file, final_plot_log, device = "pdf")
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78 }