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Int. J. Mol. Sci. 2014, 15(3), 4189-4200; doi:10.3390/ijms15034189
Article

Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage

1,2,†
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1,†
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 and
1,*
1 Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China 2 Department of Bioscience, Bengbu Medical College, Bengbu 233000, China 3 The Siberian Tiger Park of Heilongjiang, Harbin 150028, China 4 College of Wildlife Resource, Northeast Forestry University, Harbin 150028, China These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 29 October 2013 / Revised: 2 December 2013 / Accepted: 18 December 2013 / Published: 7 March 2014
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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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 Siberian tiger; Panthera tigris altaica; BAC library; genomic research; microsatellite marker
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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Liu, C.; Bai, C.; Guo, Y.; Liu, D.; Lu, T.; Li, X.; Ma, J.; Ma, Y.; Guan, W. Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage. Int. J. Mol. Sci. 2014, 15, 4189-4200.

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