Journal
BIOTECHNIQUES
Volume 53, Issue 6, Pages 365-+Publisher
FUTURE SCI LTD
DOI: 10.2144/000113962
Keywords
Pacific Biosciences; PacBio; next-generation DNA Sequencing; single molecule real time; library prep
Funding
- Wellcome Trust [098051]
- Cambridge Cancer Center
- Medical Research Council [G0801156] Funding Source: researchfish
- MRC [G0801156] Funding Source: UKRI
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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.
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