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- Title
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
- 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.
- Subjects
BACTERIAL chromosomes; ANIMAL genome mapping; SIBERIAN tiger; FIBROBLASTS; MOLECULAR cloning; GENETIC markers; GENETICS
- Publication
International Journal of Molecular Sciences, 2014, Vol 15, Issue 3, p4189
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms15034189