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Construction and characterization of a bovine BAC library with four genome-equivalent coverage

机译:具有四个基因组等效覆盖率的牛BAC文库的构建和表征

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摘要

A bovine artificial chromosome (BAC) library of 105 984 clones has been constructed in the vector pBeloBAC11 and organized in 3-dimension pools and high density membranes for screening by PCR and hybridization. The average insert size, determined after analysis of 388 clones, was estimated at 120 kb corresponding to a four genome coverage. Given the fact that a male was used to construct the library, the probability of finding any given autosomal and X or Y locus is respectively 0.98 and 0.86. The library was screened for 164 microsatellite markers and an average of 3.9 superpools was positive for each PCR system. None of the 50 or so BAC clones analysed by FISH was chimeric. This BAC library increases the international genome coverage for cattle to around 28 genome equivalents and extends the coverage of the ruminant genomes available at the Inra resource center to 15 genome equivalents.
机译:已在载体pBeloBAC11中构建了105 984个克隆的牛人工染色体(BAC)文库,并将其组织在3维池和高密度膜中,以进行PCR和杂交筛选。在分析388个克隆后确定的平均插入片段大小估计为120 kb,对应于四个基因组覆盖范围。考虑到使用男性来构建文库的事实,发现任何给定的常染色体和X或Y位点的概率分别为0.98和0.86。为该文库筛选了164个微卫星标记,每个PCR系统平均有3.9个超级池阳性。通过FISH分析的50个左右BAC克隆中,没有一个是嵌合的。该BAC库将牛的国际基因组覆盖率提高到了约28个基因组当量,并将Inra资源中心可用的反刍动物基因组的范围扩展到了15个基因组当量。

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