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About The Authors

Kengo Kato
Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Japan

Masanori Hashino
Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Japan

Tamaki Ito
Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Japan

Mari Matsui
Department of Bacteriology II, National Institute of Infectious Diseases
Japan

Satowa Suzuki
Department of Bacteriology II, National Institute of Infectious Diseases
Japan

Kumiko Kai
Department of Bacteriology II, National Institute of Infectious Diseases
Japan

Miwako Kitazume
Tomy Digital Biology Co.,ltd.
Japan

Tsuyoshi Sekizuka
Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Japan

Makoto Kuroda
Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Japan

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Home > Vol 4, No 3 (2017) > Kato

DOI: http://dx.doi.org/10.14440/jbm.2017.169

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Rapid and affordable size-selected PacBio single-molecule real-time sequencing template library construction using the bead-beating DNA extraction method

Kengo Kato, Masanori Hashino, Tamaki Ito, Mari Matsui, Satowa Suzuki, Kumiko Kai, Miwako Kitazume, Tsuyoshi Sekizuka, Makoto Kuroda

Abstract

This study demonstrated that bead-beating method facilitates a simple and rapid protocol for genomic DNA isolation for Pacific BioSciences (PacBio) sequencing with library construction of sufficient length. The protocol may also be beneficial for inactivating pathogens by simultaneous and instant DNA fragmentation, with no special equipment required to obtain large DNA fragments. This protocol was comparable in terms of quality to the standard protocol suggested by PacBioand represents an alternative, rapid shortcut for performing accurate PacBio sequencing.

Graphical Abstract

graphrical_abstract

Keywords

bead-beating, library preparation, PacBio, SMRT sequencing
Citation: Kato K, Hashino M, Ito T, Matsui M, Suzuki S, Kai K, Kitazume M, Sekizuka T, Kuroda M. Rapid and affordable size-selected PacBio single-molecule real-time sequencing template library construction using the bead-beating DNA extraction method. J Biol Methods 2017;4(3):e79. doi: 10.14440/jbm.2017.169


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