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A BAC-based STS-content map spanning a 35-Mb region of human chromosome 1p35-p36.

机译:一个基于BAC的STS内容图,跨越人类1p35-p36染色体的35-Mb区域。

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We have devised a mapping method for rapid assembly and ordering of bacterial artificial chromosome (BAC) clones on a radiation hybrid (RH) panel, using sequence-tagged sites (STSs) and PCR. The protocol consists of two rounds of two-dimensional screening from a limited number of BACs to correspond each to an STS. In the first round, STSs are assembled in the RH bins and ordered according to PCR signals derived from 384-well microtiter plates (MTPs) in which BAC clones have been arrayed. In the second round, individual BAC clones are isolated from the MTPs to build a contig. We applied this method to a 35-Mb region spanning human chromosome 1p35-p36 and assembled 1366 BACs in 11 contigs, the longest being about 20 Mb. The working draft sequences of the human genome have been integrated into the contigs to validate the accuracy. Copyright 2001 Academic Press.
机译:我们设计了一种映射方法,用于使用序列标记位点(STS)和PCR在辐射杂交(RH)面板上快速组装和排序细菌人工染色体(BAC)克隆。该协议包括从有限数量的BAC进行的两轮二维筛选,以使其分别对应于STS。在第一轮中,将STS组装在RH箱中,并根据从已排列BAC克隆的384孔微量滴定板(MTP)提取的PCR信号进行排序。在第二轮中,从MTP中分离出单个BAC克隆以构建重叠群。我们将此方法应用于跨越人类1p35-p36染色体的35 Mb区域,并在11个重叠群中组装了1366 BAC,最长的时间约为20 Mb。人类基因组的工作草图序列已整合到重叠群中以验证准确性。版权所有2001学术出版社。

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