Direct sequencing of small genomes on the Pacific Biosciences RS without library preparation. (December 2012)

• Record Type: Journal Article
• Authors: Coupland, Paul; Chandra, Tamir; Quail, Mike; Reik, Wolf; Swerdlow, Harold
• Is Part Of: Biotechniques. Volume 53:Number 6(2012)
• Page Start: 365
• Page End: 372
• Publication Date: 2012-12
• Subjects: Pacific Biosciences -- PacBio -- next-generation DNA Sequencing -- single molecule real time -- library prep; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000113962
• Languages: English
• ISSNs: 0736-6205

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.