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

Kengo Kato¹, Masanori Hashino¹, Tamaki Ito¹, Mari Matsu², Satowa Suzuki², Kumiko Kai², Miwako Kitazume³, Tsuyoshi Sekizuka¹, Makoto Kuroda¹

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¹ Laboratory of Bacterial Genomics, Pathogen Genomics Center, National Institute of Infectious Diseases, 1-23-1 Shinjyuku-ku, Tokyo 162-8640, Japan

² Department of Bacteriology II, National Institute of Infectious Diseases, Musashimurayama, Tokyo 208-0011, Japan

³ Tomy Digital Biology Co., ltd., 3-14-17 Tagara, Nerima-ku, Tokyo 179-0073, Japan

JBM 2017 , 4(3), 1;

Published: 13 September 2017

© 2017 by the author. Licensee POL Scientific, USA. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

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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.

Keywords

bead-beating

library preparation

PacBio

SMRT sequencing

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