Data production workflow for RNA-Seq. RNA-Seq requires building libraries of fragmented RNA that are then converted to cDNA by reverse transcription, followed by adaptor ligation and size selection. Sequencing libraries are prepared for clustering on an 8 lane flow cell and sequencing-by-synthesis is used to generate tens of millions of sequences per sample that can be mapped to a reference genome. The number of reads that map to a scaled region of genome space are the index of the expression level of the gene.
Malone and Oliver BMC Biology 2011 9:34 doi:10.1186/1741-7007-9-34