Difference between revisions of "Category:ECO:0000295 ! RNA-seq evidence"

name: RNA-seq evidence
def: "A type of transcript evidence based on high-throughput (HT) sequencing of fragmented cDNA molecules." [ECO:MCC]
synonym: "RNA sequencing" RELATED []
synonym: "whole transcriptome shotgun sequencing" EXACT [PMID:18611170]
synonym: "WTSS" EXACT []
is_a: ECO:0000009 ! transcript expression evidence
comment: Total RNA is isolated from cells and mRNA is purified by targeting the 3' polyadenylated (poly(A)) tail with poly(T) oligos covalently attached to a substrate. Next, cDNA fragments are generated either by hydrolysis of RNA into 200-300 base oligonucleotides, preceded by reverse transcription, or by reverse transcription initiated by random primers. The cDNA second strand is synthesized, fragments are adapter ligated and high-throughput sequencing is performed by Roche 454, Illumina, ABI-SOLiD, or another HT sequencing technology. Resulting sort reads are aligned against a reference genome, and downstream analysis is performed.