RNA-Seq

Summary of RNA-Seq. In the organism, genes are transcribed and (in a eukaryotic organism) spliced to produce mature mRNA transcripts (red). The mRNA is got from the organism, fragmented and reverse-transcribed into stable double-stranded (ds) cDNA (blue). The ds-cDNA is sequenced using high-throughput, short-read sequencing methods. These sequences can then be compared to a reference genome sequence to see which genome regions were being transcribed. This data shows where expressed genes are, their relative expression levels, and any alternative splice variants.

RNA-Seq (short for "RNA sequencing") is a technique to get snapshots of the continuously changing RNA landscape in a cell.

RNA is a nucleic acid with roles in when, where, and by how much genes are turned on. Classically, sections of DNA are copied to RNA which are decoded into proteins that carry out cellular functions.

RNAs also have many roles that fall outside this framework. RNA-Seq is typically used to analyze the amount of each gene's RNA in experimental samples. For example, in gene expression and changes made during RNA processing (alternative splicing, editing, mutations, or fusions between RNAs). RNA-Seq requires molecular biology and computational steps.

Recent advances in RNA-Seq include the ability to study single cells and entire single RNA molecules. It has broad applications in the life sciences from agriculture to medicine.

RNA-Seq Media

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  Typical RNA-Seq experimental workflow. RNA are isolated from multiple samples, converted to cDNA libraries, sequenced into a computer-readable format, aligned to a reference, and quantified for downstream analyses such as differential expression and alternative splicing. Overview of a typical RNA-Seq experimental workflow.

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  RNASeqPics

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  Typical single-cell RNA-Seq workflow. Single cells are isolated from a sample into either wells or droplets, cDNA libraries are generated and amplified, libraries are sequenced, and expression matrices are generated for downstream analyses like cell type identification.