Abstract:
:The sequencing of the full transcriptome (RNA-seq) has become the preferred choice for the measurement of genome-wide gene expression. Despite its widespread use, challenges remain in RNA-seq data analysis. One often-overlooked aspect is normalization. Despite the fact that a variety of factors or 'batch effects' can contribute unwanted variation to the data, commonly used RNA-seq normalization methods only correct for sequencing depth. The study of gene expression is particularly problematic when it is influenced simultaneously by a variety of biological factors in addition to the one of interest. Using examples from experimental neuroscience, we show that batch effects can dominate the signal of interest; and that the choice of normalization method affects the power and reproducibility of the results. While commonly used global normalization methods are not able to adequately normalize the data, more recently developed RNA-seq normalization can. We focus on one particular method, RUVSeq and show that it is able to increase power and biological insight of the results. Finally, we provide a tutorial outlining the implementation of RUVSeq normalization that is applicable to a broad range of studies as well as meta-analysis of publicly available data.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Peixoto L,Risso D,Poplawski SG,Wimmer ME,Speed TP,Wood MA,Abel Tdoi
10.1093/nar/gkv736subject
Has Abstractpub_date
2015-09-18 00:00:00pages
7664-74issue
16eissn
0305-1048issn
1362-4962pii
gkv736journal_volume
43pub_type
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