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Abstract
<p class="first" id="P1">Deep sequencing has been revolutionizing biology and medicine
in recent years, providing
single base-level precision for our understanding of nucleic acid sequences in high
throughput fashion. Sequencing of RNA, or RNA-Seq, is now a common method to analyze
gene expression and to uncover novel RNA species. Aspects of RNA biogenesis and metabolism
can be interrogated with specialized methods for cDNA library preparation. In this
study, we review current RNA-Seq methods for general analysis of gene expression and
several specific applications, including isoform and gene fusion detection, digital
gene expression profiling, targeted sequencing and single-cell analysis. In addition,
we discuss approaches to examine aspects of RNA in the cell, technical challenges
of existing RNA-Seq methods, and future directions.
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