title: Direct sequencing of small genomes on the Pacific Biosciences RS without library preparation.
abstract: We have developed a sequencing method on the Pacific Biosciences RS sequencer (the PacBio) for small DNA molecules that avoids the need for a standard library preparation. To date this approach has been applied toward sequencing single-stranded and double-stranded viral genomes, bacterial plasmids, plasmid vector models for DNA-modification analysis, and linear DNA fragments covering an entire bacterial genome. Using direct sequencing it is possible to generate sequence data from as little as 1 ng of DNA, offering a significant advantage over current protocols which typically require 400-500 ng of sheared DNA for the library preparation.
authors.affiliation: Wellcome Trust Sanger Institute, Hinxton, UK. pc10@sanger.ac.uk
authors.forename: Paul
authors.initials: P
authors.lastname: Coupland
authors.forename: Tamir
authors.initials: T
authors.lastname: Chandra
authors.forename: Mike
authors.initials: M
authors.lastname: Quail
authors.forename: Wolf
authors.initials: W
authors.lastname: Reik
authors.forename: Harold
authors.initials: H
authors.lastname: Swerdlow
mesh_terms: D001483:Base Sequence; D015723:Gene Library; D016680:Genome, Bacterial; D023281:Genomics / Q000379:methods; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods*; D012984:Software; D014584:User-Computer Interface