RACE-sequencing (RACE-seq) refers to a method in molecular biology for characterizing cDNA molecules generated by Rapid amplification of cDNA ends using high-throughput sequencing technologies.
High-throughput sequencing characterization of RACE fragments is highly time-efficient, more sensitive, less costly and technically feasible compared to traditional characterization of RACE fragments with molecular cloning followed by Sanger sequencing of a few clones.
History and applications
RACE can be used to amplify unknown 5' (5'-RACE) or 3' (3'-RACE) parts of RNA molecules where part of the RNA sequence is known and targeted by a gene-specific primer. Combined with high-throughput sequencing for characterization of these amplified RACE products, it is possible to apply the approach to characterize any types of coding or non-coding RNA-molecules.
The idea of combining RACE with high-throughput sequencing was first introduced in 2009 as Deep-RACE to perform mapping of Transcription start sites (TSS) of 17 genes in a single cell-line. In a study from 2014 to accurately map cleavage sites of target RNA directed by synthetic siRNAs, the approach was first named RACE-seq. Further, the methodology was used to characterize full-length unknown parts of novel transcripts and fusion transcripts in colorectal cancer. In another study aiming to characterize unknown transcript structures of lncRNAs, RACE was used in combination with semi-long 454 sequencing