De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis.

De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis.
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DOI:
10.1038/nprot.2013.084
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发表时间:
2013-08
期刊:
影响因子:
14.8
通讯作者:
--
中科院分区:
生物学1区
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--
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RNA-Seq数据的从头组装使我们能够在不需要基因组序列的情况下研究转录组,例如在具有生态和进化重要性的非模式生物、癌症样本或微生物组中。在本方案中,我们描述了使用Trinity平台从非模型生物体中的RNA-Seq数据进行从头转录组组装。我们还介绍了Trinity支持的下游应用配套工具,包括用于转录丰度估计的RSEM,用于识别样品中差异表达转录本的R/Bioconductor软件包,以及识别蛋白质编码基因的方法。在包含的教程中,我们提供了利用Trinity平台进行基因组独立转录组分析的工作流程。软件、文档和演示可从http://trinityrnaseq.sf.net免费获得。
De novo assembly of RNA-Seq data allows us to study transcriptomes without the need for a genome sequence, such as in non-model organisms of ecological and evolutionary importance, cancer samples, or the microbiome. In this protocol, we describe the use of the Trinity platform for de novo transcriptome assembly from RNA-Seq data in non-model organisms. We also present Trinity’s supported companion utilities for downstream applications, including RSEM for transcript abundance estimation, R/Bioconductor packages for identifying differentially expressed transcripts across samples, and approaches to identify protein coding genes. In an included tutorial we provide a workflow for genome-independent transcriptome analysis leveraging the Trinity platform. The software, documentation and demonstrations are freely available from http://trinityrnaseq.sf.net.
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