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

Authors :
Kae Koganebuchi
Takashi Gakuhari
Hirohiko Takeshima
Kimitoshi Sato
Kiyotaka Fujii
Toshihiro Kumabe
Satoshi Kasagi
Takehiro Sato
Atsushi Tajima
Hiroki Shibata
Motoyuki Ogawa
Hiroki Oota
Source :
PLoS ONE, Vol 13, Iss 7, p e0200170 (2018)
Publication Year :
2018
Publisher :
Public Library of Science (PLoS), 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.

Subjects

Subjects :
Medicine
Science

Details

Language :
English
ISSN :
19326203
Volume :
13
Issue :
7
Database :
Directory of Open Access Journals
Journal :
PLoS ONE
Publication Type :
Academic Journal
Accession number :
edsdoj.7f40fbf1068f41f4af4cb32340962b0d
Document Type :
article
Full Text :
https://doi.org/10.1371/journal.pone.0200170