Reproducibility of high-throughput mRNA and small RNA sequencing across laboratories
作者： Peter AC't Hoen , Marc R Friedländer , Jonas Almlöf , Michael Sammeth , Irina Pulyakhina
DOI: 10.1038/NBT.2702
关键词:
摘要: RNA sequencing is an increasingly popular technology for genome-wide analysis of transcript sequence and abundance. However, understanding the sources technical interlaboratory variation still limited. To address this, GEUVADIS consortium sequenced mRNAs small RNAs lymphoblastoid cell lines 465 individuals in seven centers, with a large number replicates. The between laboratories appeared to be considerably smaller than already limited biological variation. Laboratory effects were mainly seen differences insert size GC content could adequately corrected for. In small-RNA sequencing, microRNA (miRNA) differed widely samples owing competitive rRNA fragments. This did not affect relative quantification miRNAs. We conclude that distributing among different feasible, given proper standardization randomization procedures. provide set quality measures guidelines assessing biases RNA-seq data.
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