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Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage
Authors:Changqing Liu  Chunyu Bai  Yu Guo  Dan Liu  Taofeng Lu  Xiangchen Li  Jianzhang Ma  Yuehui Ma  Weijun Guan
Affiliation:1.Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; E-Mails: (C.L.); (C.B.); (T.L.); (X.L.); (Y.M.);2.Department of Bioscience, Bengbu Medical College, Bengbu 233000, China; E-Mail: ;3.The Siberian Tiger Park of Heilongjiang, Harbin 150028, China; E-Mail: ;4.College of Wildlife Resource, Northeast Forestry University, Harbin 150028, China; E-Mail:
Abstract:Bacterial artificial chromosome (BAC) libraries are extremely valuable for the genome-wide genetic dissection of complex organisms. The Siberian tiger, one of the most well-known wild primitive carnivores in China, is an endangered animal. In order to promote research on its genome, a high-redundancy BAC library of the Siberian tiger was constructed and characterized. The library is divided into two sub-libraries prepared from blood cells and two sub-libraries prepared from fibroblasts. This BAC library contains 153,600 individually archived clones; for PCR-based screening of the library, BACs were placed into 40 superpools of 10 × 384-deep well microplates. The average insert size of BAC clones was estimated to be 116.5 kb, representing approximately 6.46 genome equivalents of the haploid genome and affording a 98.86% statistical probability of obtaining at least one clone containing a unique DNA sequence. Screening the library with 19 microsatellite markers and a SRY sequence revealed that each of these markers were present in the library; the average number of positive clones per marker was 6.74 (range 2 to 12), consistent with 6.46 coverage of the tiger genome. Additionally, we identified 72 microsatellite markers that could potentially be used as genetic markers. This BAC library will serve as a valuable resource for physical mapping, comparative genomic study and large-scale genome sequencing in the tiger.
Keywords:Siberian tiger  Panthera tigris altaica  BAC library  genomic research  microsatellite marker
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