July 7, 2019  |  

Direct sequencing of small genomes on the Pacific Biosciences RS without library preparation.

Authors: Coupland, Paul and Chandra, Tamir and Quail, Mike and Reik, Wolf and Swerdlow, Harold

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.

Journal: BioTechniques
DOI: 10.2144/000113962
Year: 2012

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