Abstract：Eukaryotic transcriptome is diverse and complex. RNA sequencing (RNA-seq) can be used to provide a global profile of the transcriptome, find biomarkers associated with dieases and support the diagnosis and treatment of complex diseases. The process of RNA-seq includes sample preparation, library preparation, sequencing and analysis. The technical errors that are introduced during all the workflow influence the accuracy of RNA-seq. These errors can be understood and mitigated through the use of RNA spike-in, which can determine quality control for RNA-seq and improve the reliability of RNA-seq data. Therefore, RNA spike-in and its applications are introduced, including normalization between samples, gene expression measurements, detection and quantification of diverse transcripts, which provides powerful reference for improving the quality of RNA-seq by using RNA spike-in effectively.
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