Supporting data for "A workflow for simplified analysis of ATAC-cap-seq data in R"
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ATAC-cap-seq is a high-throughput sequencing method that combines targeted nucleic acid enrichment of precipitated DNA fragments with an upstream ATAC-seq step. There are increased analytical difficulties arising from working with a set of regions of interest that may be small in number and biologically dependent. Common statistical pipelines for RNAseq might be assumed to apply but can give misleading results on ATAC-cap-seq data. A tool is needed to allow a non-specialist user to quickly and easily summarise data and apply sensible and effective normalisation and analysis. We developed atacR to allow a user to easily analyse their ATAC enrichment experiment. It provides comprehensive summary functions and diagnostic plots for studying enriched tag abundance. Applying between-sample normalisation is made straightforward and functions for normalising based on user-defined control regions, whole library size and regions selected from the least variable regions in a dataset are provided. Three methods for detecting differential abundance of tags from enriched methods are provided, including Bootstrap t, Bayes Factor and a wrapped version of the standard exact test in the edgeR package. We compared the precision, recall and F-score of each detection method on resampled datasets at varying replicate, significance threshold and genes changed, we found that the Bayes factor method had greatest overall detection power, though edgeR was slightly stronger in simulations with lower numbers of genes changed. Our package allows a non-specialist user to easily and effectively apply methods appropriate to the analysis of ATAC-cap-seq in a reproducible manner. The package is implemented in pure R and is fully interoperable with common workflows in Bioconductor.



