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

doi:10.14440/jbm.2017.169

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

Authors

Kengo Kato Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Masanori Hashino Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Tamaki Ito Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Mari Matsui Department of Bacteriology II, National Institute of Infectious Diseases
Satowa Suzuki Department of Bacteriology II, National Institute of Infectious Diseases
Kumiko Kai Department of Bacteriology II, National Institute of Infectious Diseases
Miwako Kitazume Tomy Digital Biology Co.,ltd.
Tsuyoshi Sekizuka Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.
Makoto Kuroda Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases.

DOI:

https://doi.org/10.14440/jbm.2017.169

Keywords:

bead-beating, library preparation, PacBio, SMRT sequencing

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.

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References

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Published

2017-09-13

How to Cite

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 [Internet]. 2017Sep.13 [cited 2022Oct.6];4(3):e79. Available from: https://jbmethods.org/jbm/article/view/169

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Vol. 4 No. 3 (2017)

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Articles

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