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      Understanding sequencing data as compositions: an outlook and review

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          Abstract

          Motivation

          Although seldom acknowledged explicitly, count data generated by sequencing platforms exist as compositions for which the abundance of each component (e.g. gene or transcript) is only coherently interpretable relative to other components within that sample. This property arises from the assay technology itself, whereby the number of counts recorded for each sample is constrained by an arbitrary total sum (i.e. library size). Consequently, sequencing data, as compositional data, exist in a non-Euclidean space that, without normalization or transformation, renders invalid many conventional analyses, including distance measures, correlation coefficients and multivariate statistical models.

          Results

          The purpose of this review is to summarize the principles of compositional data analysis (CoDA), provide evidence for why sequencing data are compositional, discuss compositionally valid methods available for analyzing sequencing data, and highlight future directions with regard to this field of study.

          Supplementary information

          Supplementary data are available at Bioinformatics online.

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          Most cited references32

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          Data quality aware analysis of differential expression in RNA-seq with NOISeq R/Bioc package

          As the use of RNA-seq has popularized, there is an increasing consciousness of the importance of experimental design, bias removal, accurate quantification and control of false positives for proper data analysis. We introduce the NOISeq R-package for quality control and analysis of count data. We show how the available diagnostic tools can be used to monitor quality issues, make pre-processing decisions and improve analysis. We demonstrate that the non-parametric NOISeqBIO efficiently controls false discoveries in experiments with biological replication and outperforms state-of-the-art methods. NOISeq is a comprehensive resource that meets current needs for robust data-aware analysis of RNA-seq differential expression.
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            Synthetic spike-in standards for RNA-seq experiments.

            High-throughput sequencing of cDNA (RNA-seq) is a widely deployed transcriptome profiling and annotation technique, but questions about the performance of different protocols and platforms remain. We used a newly developed pool of 96 synthetic RNAs with various lengths, and GC content covering a 2(20) concentration range as spike-in controls to measure sensitivity, accuracy, and biases in RNA-seq experiments as well as to derive standard curves for quantifying the abundance of transcripts. We observed linearity between read density and RNA input over the entire detection range and excellent agreement between replicates, but we observed significantly larger imprecision than expected under pure Poisson sampling errors. We use the control RNAs to directly measure reproducible protocol-dependent biases due to GC content and transcript length as well as stereotypic heterogeneity in coverage across transcripts correlated with position relative to RNA termini and priming sequence bias. These effects lead to biased quantification for short transcripts and individual exons, which is a serious problem for measurements of isoform abundances, but that can partially be corrected using appropriate models of bias. By using the control RNAs, we derive limits for the discovery and detection of rare transcripts in RNA-seq experiments. By using data collected as part of the model organism and human Encyclopedia of DNA Elements projects (ENCODE and modENCODE), we demonstrate that external RNA controls are a useful resource for evaluating sensitivity and accuracy of RNA-seq experiments for transcriptome discovery and quantification. These quality metrics facilitate comparable analysis across different samples, protocols, and platforms.
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              Revisiting global gene expression analysis.

              Gene expression analysis is a widely used and powerful method for investigating the transcriptional behavior of biological systems, for classifying cell states in disease, and for many other purposes. Recent studies indicate that common assumptions currently embedded in experimental and analytical practices can lead to misinterpretation of global gene expression data. We discuss these assumptions and describe solutions that should minimize erroneous interpretation of gene expression data from multiple analysis platforms. Copyright © 2012 Elsevier Inc. All rights reserved.
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                Author and article information

                Contributors
                Role: Associate Editor
                Journal
                Bioinformatics
                Bioinformatics
                bioinformatics
                Bioinformatics
                Oxford University Press
                1367-4803
                1367-4811
                15 August 2018
                28 March 2018
                28 March 2018
                : 34
                : 16
                : 2870-2878
                Affiliations
                [1 ]Bioinformatics Core Research Group, Deakin University, Geelong, Australia
                [2 ]Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Barcelona, Spain
                [3 ]Universitat Pompeu Fabra (UPF), Barcelona, Spain
                [4 ]Centre for Integrative Ecology, School of Life and Environmental Sciences, Deakin University, Geelong, Australia
                [5 ]Poultry Hub Australia, University of New England, Armidale, Australia
                Author notes
                To whom correspondence should be addressed. Email: contacttomquinn@ 123456gmail.com
                Author information
                http://orcid.org/0000-0003-0286-6329
                Article
                bty175
                10.1093/bioinformatics/bty175
                6084572
                29608657
                91971681-8b42-4fc9-b6c2-ba1119184dc2
                © The Author(s) 2018. Published by Oxford University Press.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

                History
                : 24 October 2017
                : 20 March 2018
                : 26 March 2018
                Page count
                Pages: 9
                Categories
                Review
                Gene Expression

                Bioinformatics & Computational biology
                Bioinformatics & Computational biology

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