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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Gene Library
  • Genome, Bacterial
  • Genomics / methods
  • Molecular Sequence Data
  • Sequence Analysis, DNA / instrumentation*
  • Sequence Analysis, DNA / methods*
  • Software*
  • User-Computer Interface