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RNA-seq differential expression studies: more sequence or more replication?


ABSTRACT: MOTIVATION:RNA-seq is replacing microarrays as the primary tool for gene expression studies. Many RNA-seq studies have used insufficient biological replicates, resulting in low statistical power and inefficient use of sequencing resources. RESULTS:We show the explicit trade-off between more biological replicates and deeper sequencing in increasing power to detect differentially expressed (DE) genes. In the human cell line MCF7, adding more sequencing depth after 10 M reads gives diminishing returns on power to detect DE genes, whereas adding biological replicates improves power significantly regardless of sequencing depth. We also propose a cost-effectiveness metric for guiding the design of large-scale RNA-seq DE studies. Our analysis showed that sequencing less reads and performing more biological replication is an effective strategy to increase power and accuracy in large-scale differential expression RNA-seq studies, and provided new insights into efficient experiment design of RNA-seq studies. AVAILABILITY AND IMPLEMENTATION:The code used in this paper is provided on: http://home.uchicago.edu/?jiezhou/replication/. The expression data is deposited in the Gene Expression Omnibus under the accession ID GSE51403.

SUBMITTER: Liu Y 

PROVIDER: S-EPMC3904521 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature

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RNA-seq differential expression studies: more sequence or more replication?

Liu Yuwen Y   Zhou Jie J   White Kevin P KP  

Bioinformatics (Oxford, England) 20131206 3


<h4>Motivation</h4>RNA-seq is replacing microarrays as the primary tool for gene expression studies. Many RNA-seq studies have used insufficient biological replicates, resulting in low statistical power and inefficient use of sequencing resources.<h4>Results</h4>We show the explicit trade-off between more biological replicates and deeper sequencing in increasing power to detect differentially expressed (DE) genes. In the human cell line MCF7, adding more sequencing depth after 10 M reads gives d  ...[more]

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