Dataset Information

The Selection of Quantification Pipelines for Illumina RNA-seq Data Using a Subsampling Approach.

ABSTRACT: RNA sequencing, or (RNA-seq for short,, is a widely applied technology that for extractings gene and transcript expression from biological samples. Given numerous quantification pipelines for RNA-seq data, one fundamental challenge is to determine identify a pipeline that can produce the most accurate estimate the most accurate gene and/or transcript expression. Exploring all available pipelines requires tremendous extensive computational resources, so. Therefore, we propose to use a subsampling approach that can improve speed up the pipeline evaluation and selection the efficiency process of pipeline performance evaluation for a given RNA-seq dataset. We applied our approach to one simulated and two real RNA-seq datasets and found that expression estimates derived from subsampled data are close surrogates for those derived from original data. In addition, the ranking of quantification pipelines based on the subsampled data was highly correlated concordant with that based on the original data. Therefore, we conclude that subsampling is a valid approach to facilitating efficient quantification pipeline selection using RNA-seq data.

SUBMITTER: Wu PY

PROVIDER: S-EPMC5267345 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature

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The Selection of Quantification Pipelines for Illumina RNA-seq Data Using a Subsampling Approach.

Wu Po-Yen PY Wang May D MD

... IEEE-EMBS International Conference on Biomedical and Health Informatics. IEEE-EMBS International Conference on Biomedical and Health Informatics 20160201

RNA sequencing, or (RNA-seq for short,, is a widely applied technology that for extractings gene and transcript expression from biological samples. Given numerous quantification pipelines for RNA-seq data, one fundamental challenge is to determine identify a pipeline that can produce the most accurate estimate the most accurate gene and/or transcript expression. Exploring all available pipelines requires tremendous extensive computational resources, so. Therefore, we propose to use a subsampling ...[more]

PMID: 28133637

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Project description:MotivationDespite numerous RNA-seq samples available at large databases, most RNA-seq analysis tools are evaluated on a limited number of RNA-seq samples. This drives a need for methods to select a representative subset from all available RNA-seq samples to facilitate comprehensive, unbiased evaluation of bioinformatics tools. In sequence-based approaches for representative set selection (e.g. a k-mer counting approach that selects a subset based on k-mer similarities between RNA-seq samples), because of the large numbers of available RNA-seq samples and of k-mers/sequences in each sample, computing the full similarity matrix using k-mers/sequences for the entire set of RNA-seq samples in a large database (e.g. the SRA) has memory and runtime challenges; this makes direct representative set selection infeasible with limited computing resources.ResultsWe developed a novel computational method called 'hierarchical representative set selection' to handle this challenge. Hierarchical representative set selection is a divide-and-conquer-like algorithm that breaks representative set selection into sub-selections and hierarchically selects representative samples through multiple levels. We demonstrate that hierarchical representative set selection can achieve summarization quality close to that of direct representative set selection, while largely reducing runtime and memory requirements of computing the full similarity matrix (up to 8.4× runtime reduction and 5.35× memory reduction for 10 000 and 12 000 samples respectively that could be practically run with direct subset selection). We show that hierarchical representative set selection substantially outperforms random sampling on the entire SRA set of RNA-seq samples, making it a practical solution to representative set selection on large databases like the SRA.Availability and implementationThe code is available at https://github.com/Kingsford-Group/hierrepsetselection and https://github.com/Kingsford-Group/jellyfishsim.Supplementary informationSupplementary data are available at Bioinformatics online.

Project description:Most microbes in the natural environment are difficult to cultivate. Thus, culture-independent analysis for microbial community structure is important for the understanding of its ecological functions. An immense ribosomal RNA sequence collection is available from phylogenetic research on organisms in all domains. These sequences are available for use in genetic research. However, the amplicon-seq process using PCR requires the construction of a sequence library. Construction can introduce bias into quantitative analyses, and each domain of species needs its own primer set. Total RNA sequencing has the advantage of analyzing an entire microbial community, including bacteria, archea, and eukaryote, at once. Such analysis yields large amounts of ribosomal RNA sequences that can be used for analysis without PCR bias. Evaluation using total RNA-seq for quantitative analysis of microbial communities and comparison with amplicon-seq is still rare. In the present study, we developed a mapping-based total RNA-seq analysis to obtain quantitative information on microbial community structure and compared our results with ordinary amplicon-seq methods. We read total RNA sequences from a commercially available mock community (ATCC MSA-2003) and divided reads into small subunit ribosomal RNA (ssrRNA) origin reads and others, such as mRNA origin reads. We then mapped ssrRNA origin reads on annotated assembled contigs and obtained quantitative results under several analysis strategies. Removal of low complexity sequences, sorting ssrRNA with paired-in mode, and performing homology-based taxonomical assignments (BLAST+ or vsearch) showed superior outcomes to other strategies. Results with this approach showed a median relative abundance among ten mock community members of ~10%; ordinary amplicon-seq showed a much lower percentage. Thus, total RNA-seq can be a powerful tool for analyzing microbial community structure and is not limited to analyzing gene expression profiling of microbiomes.

Dataset Information

The Selection of Quantification Pipelines for Illumina RNA-seq Data Using a Subsampling Approach.

Publications

The Selection of Quantification Pipelines for Illumina RNA-seq Data Using a Subsampling Approach.

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