RT Journal Article
SR Electronic
T1 A new targeted capture method using bacterial artificial chromosome (BAC) libraries as baits for sequencing relatively large genes
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 302760
DO 10.1101/302760
A1 Kae Koganebuchi
A1 Takashi Gakuhari
A1 Hirohiko Takeshima
A1 Kimitoshi Sato
A1 Kiyotaka Fujii
A1 Toshihiro Kumabe
A1 Satoshi Kasagi
A1 Takehiro Sato
A1 Atsushi Tajima
A1 Hiroki Shibata
A1 Motoyuki Ogawa
A1 Hiroki Oota
YR 2018
UL http://biorxiv.org/content/early/2018/04/17/302760.abstract
AB 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 region with substantial length.