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Biotechniques. 2012 Dec;53(6):365-72. doi: 10.2144/000113962.

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

Author information

1
Wellcome Trust Sanger Institute, Hinxton, UK. pc10@sanger.ac.uk

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.

PMID:
23227987
PMCID:
PMC3808810
DOI:
10.2144/000113962
[Indexed for MEDLINE]
Free PMC Article

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