title: ACC human 2K ver6.0.
status: Public on Apr 01 2014
contact.city: Nagoya
contact.first: Miyuki
contact.middle: Suguro
contact.last: Katayama
contact.email: mkatayama@aichi-cc.jp
contact.state: Aichi
contact.address: 1-1 Kanokoden, Chikusa-ku
contact.department: Division of Molecular Medicine
contact.country: Japan
contact.institute: Aichi Cancer Center Research Institute
contact.phone: +81-52-762-6111
organism: Homo sapiens
manufacture_protocol: The array consisted of 2,400 bacterial artificial chromosome (BAC) and P-1 derived artificial chromosome (PAC) clones, covering the human genome at roughly a 1.5-Mb resolution, from library RP11 and 13 for BAC clones and RP1, 3, 4, and 5 for PAC clones. These clones were obtained from the BACPAC Resource Center at the Children’s Hospital Oakland Research Institute (Oakland, CA). 4 Each clone was cultured in Terrific Broth medium with the chloramphenicol (25 μg/ml), and BAC and PAC DNA was extracted with a plasmid Mini-kit (Qiagen). The location of each clone was also confirmed by FISH analysis. Roughly 10% of these clones could not be assigned to their expected region and excluded from this study, whereas the confirmed clones were used for array CGH. We used 10 ng of BAC and PAC DNA as the template for degenerate oligonucleotide primed PCR with the primer 5′-CCGACTCGAGNNNNNNATGTGG-3′, where n = A, C, G, or T (24) . Amplifications were performed on a TaKaRa PCR Thermal Cycler MP (TaKaRa) using ExTaq polymerase (TaKaRa). Degenerate oligonucleotide primed-PCR products were enriched by ethanol precipitation and dissolved in distilled water, and then equal volume of DNA spotting solution DSP0050 (MATSUNAMI, Osaka, Japan) was added (∼1 μg/μl). DNA was robotically spotted (NGK Insulators, Ltd.) in duplicate onto CodeLink activated slides (GE Healthcare Life Sciences).