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

J Biol Methods. 2017 Sep 13;4(3):e79. doi: 10.14440/jbm.2017.169. eCollection 2017.

Authors

Kengo Kato¹, Masanori Hashino¹, Tamaki Ito¹, Mari Matsui², Satowa Suzuki², Kumiko Kai², Miwako Kitazume³, Tsuyoshi Sekizuka¹, Makoto Kuroda¹

Affiliations

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.

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 PacBio and represents an alternative, rapid shortcut for performing accurate PacBio sequencing.

Keywords: PacBio; SMRT sequencing; bead-beating; library preparation.