Construction and characterization of a human bacterial artificial chromosome library
Construction and characterization of a human bacterial artificial chromosome library
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DOI：
10.1006/geno.1996.0268
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发表时间：
1996-06-01
期刊：
影响因子：
4.4
通讯作者：
Shizuya, H
中科院分区：
文献类型：
--
作者：
Kim, UJ;Birren, BW;Shizuya, H
我们已经构建了具有约4x覆盖度的阵列式人类基因组BAC文库，其由96，000个BAC克隆表示，平均插入大小接近140 kb。一种新的BAC载体，允许基于颜色的阳性筛选，以确定具有插入片段的转化体，提高了BAC克隆效率。将文库以高密度网格化到杂交滤膜上，以通过菌落杂交有效地鉴定BAC克隆。还将文库配制成特征性DNA池，以允许针对STS内容物对文库进行PCR筛选。我们主要通过筛选300多个不同的地标，包括cDNA，STS，和粘粒克隆库的特点。我们描述了使用BAC克隆的方法，并讨论了基因组表征，绘图和测序的影响。(C)出版社：Academic Press，Inc.
We have constructed an arrayed human genomic BAC library with approximately 4x coverage that is represented by 96,000 BAC clones with average insert size of nearly 140 kb. A new BAC vector that allows color-based positive screening to identify transformants with inserts has increased BAC cloning efficiency. The library was gridded onto hybridization filters at high density for efficient identification of BAC clones by colony hybridization. The library was also formulated into characteristic DNA pools to allow for PCR screening of the library for STS content. We have characterized the library mainly by screening with more than 300 different landmarks that include cDNA, STSs, and cosmid clones. We describe methods for using BAC clones and discuss the implications for genome characterization, mapping, and sequencing. (C) 1996 Academic Press, Inc.