RNA sequencing

RNA sequencing (RNA-seq) converts RNA into sequenceable molecules and reads them to identify RNA species and estimate their abundance in a sample.

RNA extraction: The isolation of RNA molecules from cells, tissues, or other samples. Clean RNA is the starting material for library preparation.
Messenger RNA: An RNA molecule that carries protein-coding information from DNA to ribosomes. Poly(A)-selected RNA-seq primarily measures mature messenger RNA.
Differential gene expression: The statistical comparison of gene or transcript abundance between biological conditions. This is a common use of normalized RNA-seq counts across samples.
Microarray: A hybridization-based assay that measures nucleic acid abundance using probes fixed to a surface. Unlike RNA-seq, microarrays detect only sequences represented by designed probes.
Complementary DNA: DNA synthesized from an RNA template by reverse transcriptase. Most RNA-seq platforms sequence DNA copies rather than RNA directly.
Ribosomal RNA: RNA molecules that form the structural and catalytic core of ribosomes. Ribosomal RNA dominates total cellular RNA and is often depleted before sequencing.
Single-cell RNA sequencing: RNA sequencing that measures transcripts separately in individual cells. It resolves cell-to-cell expression differences hidden in bulk samples.
Quantitative PCR: A method that quantifies selected DNA or complementary DNA targets during amplification. It measures chosen targets precisely, while RNA-seq surveys many transcripts at once.
RNA-seq library preparation: The conversion of RNA or complementary DNA into fragments prepared for sequencing. Adapters and sample indexes make RNA-derived fragments readable and traceable.
Non-coding RNA: RNA that functions without encoding a protein. RNA-seq can reveal abundant non-coding transcripts missed by protein-focused assays.

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