A new targeted capture method using bacterial artificial chromosome (BAC) libraries as baits for sequencing relatively large genes

Kae Koganebuchi, Takashi Gakuhari, Hirohiko Takeshima, Kimitoshi Sato, Kiyotaka Fujii, Toshihiro Kumabe, Satoshi Kasagi, Takehiro Sato, Atsushi Tajima, Hiroki Shibata, Motoyuki Ogawa, Hiroki Oota

Research output: Contribution to journalArticle

Abstract

To analyze a specific genome region using next-generation sequencing technologies, the enrichment of DNA libraries with targeted capture methods has been standardized. For enrichment of mitochondrial genome, a previous study developed an original targeted capture method that use baits constructed from long-range polymerase chain reaction (PCR) amplicons, common laboratory reagents, and equipment. In this study, a new targeted capture method is presented, that of bacterial artificial chromosome (BAC) double capture (BDC), modifying the previous method, but using BAC libraries as baits for sequencing a relatively large gene. We applied the BDC approach for the 214 kb autosomal region, ring finger protein 213, which is the susceptibility gene of moyamoya disease (MMD). To evaluate the reliability of BDC, cost and data quality were compared with those of a commercial kit. While the ratio of duplicate reads was higher, the cost was less than that of the commercial kit. The data quality was sufficiently the same as that of the kit. Thus, BDC can be an easy, low-cost, and useful method for analyzing individual genome regions with substantial length.

Original languageEnglish
Article numbere0200170
JournalPloS one
Volume13
Issue number7
DOIs
Publication statusPublished - Jul 2018

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Koganebuchi, K., Gakuhari, T., Takeshima, H., Sato, K., Fujii, K., Kumabe, T., Kasagi, S., Sato, T., Tajima, A., Shibata, H., Ogawa, M., & Oota, H. (2018). A new targeted capture method using bacterial artificial chromosome (BAC) libraries as baits for sequencing relatively large genes. PloS one, 13(7), [e0200170]. https://doi.org/10.1371/journal.pone.0200170