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

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-redundanc...

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Autores principales: Liu, Changqing, Bai, Chunyu, Guo, Yu, Liu, Dan, Lu, Taofeng, Li, Xiangchen, Ma, Jianzhang, Ma, Yuehui, Guan, Weijun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3975391/
https://www.ncbi.nlm.nih.gov/pubmed/24608928
http://dx.doi.org/10.3390/ijms15034189
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author Liu, Changqing
Bai, Chunyu
Guo, Yu
Liu, Dan
Lu, Taofeng
Li, Xiangchen
Ma, Jianzhang
Ma, Yuehui
Guan, Weijun
author_facet Liu, Changqing
Bai, Chunyu
Guo, Yu
Liu, Dan
Lu, Taofeng
Li, Xiangchen
Ma, Jianzhang
Ma, Yuehui
Guan, Weijun
author_sort Liu, Changqing
collection PubMed
description 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.
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spelling pubmed-39753912014-04-04 Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage Liu, Changqing Bai, Chunyu Guo, Yu Liu, Dan Lu, Taofeng Li, Xiangchen Ma, Jianzhang Ma, Yuehui Guan, Weijun Int J Mol Sci Article 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. Molecular Diversity Preservation International (MDPI) 2014-03-07 /pmc/articles/PMC3975391/ /pubmed/24608928 http://dx.doi.org/10.3390/ijms15034189 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland http://creativecommons.org/licenses/by/3.0/ This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Liu, Changqing
Bai, Chunyu
Guo, Yu
Liu, Dan
Lu, Taofeng
Li, Xiangchen
Ma, Jianzhang
Ma, Yuehui
Guan, Weijun
Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage
title Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage
title_full Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage
title_fullStr Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage
title_full_unstemmed Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage
title_short Construction and Analysis of Siberian Tiger Bacterial Artificial Chromosome Library with Approximately 6.5-Fold Genome Equivalent Coverage
title_sort construction and analysis of siberian tiger bacterial artificial chromosome library with approximately 6.5-fold genome equivalent coverage
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3975391/
https://www.ncbi.nlm.nih.gov/pubmed/24608928
http://dx.doi.org/10.3390/ijms15034189
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