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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.

SUBMITTER: Coupland P 

PROVIDER: S-EPMC3808810 | biostudies-literature | 2012 Dec

REPOSITORIES: biostudies-literature

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

Coupland Paul P   Chandra Tamir T   Quail Mike M   Reik Wolf W   Swerdlow Harold H  

BioTechniques 20121201 6


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  ...[more]

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