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

PLoS One. 2018 Jul 12;13(7):e0200170. doi: 10.1371/journal.pone.0200170. eCollection 2018.

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adenosine Triphosphatases / genetics
  • Chromosomes, Artificial, Bacterial / genetics*
  • Gene Library*
  • Genetic Predisposition to Disease
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Moyamoya Disease / genetics
  • Sequence Analysis, DNA / methods*
  • Ubiquitin-Protein Ligases / genetics

Substances

  • RNF213 protein, human
  • Ubiquitin-Protein Ligases
  • Adenosine Triphosphatases

Grants and funding

This work was supported by KAKENHI (https://www.jsps.go.jp/j-grantsinaid/, Grants-in-Aid for Scientific Research B and A) to HO (25284157 and 18H03593, respectively) from the Japan Society for the Promotion of Science (JSPS). KK was supported by the JSPS Fellowship (https://www.jsps.go.jp/j-pd/, 15J10824). This work was partly performed in the Cooperative Research Project Program of the Medical Institute of Bioregulation, Kyushu University (http://www.bioreg.kyushu-u.ac.jp/mib/activities_collabo_j.html). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.