title: Library preparation for highly accurate population sequencing of RNA viruses.
abstract: Circular resequencing (CirSeq) is a novel technique for efficient and highly accurate next-generation sequencing (NGS) of RNA virus populations. The foundation of this approach is the circularization of fragmented viral RNAs, which are then redundantly encoded into tandem repeats by 'rolling-circle' reverse transcription. When sequenced, the redundant copies within each read are aligned to derive a consensus sequence of their initial RNA template. This process yields sequencing data with error rates far below the variant frequencies observed for RNA viruses, facilitating ultra-rare variant detection and accurate measurement of low-frequency variants. Although library preparation takes ∼5 d, the high-quality data generated by CirSeq simplifies downstream data analysis, making this approach substantially more tractable for experimentalists.
authors: Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, USA.
authors: Ashley
authors: A
authors: Acevedo
authors: Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, USA.
authors: Raul
authors: R
authors: Andino
mesh_terms: D000465:Algorithms; D015723:Gene Library; D014644:Genetic Variation; D012328:RNA Viruses / Q000235:genetics*; D012367:RNA, Viral / Q000737:chemistry*; D017423:Sequence Analysis, RNA / Q000379:methods*; D020080:Tandem Repeat Sequences
chemical_list: D012367:RNA, Viral