一个基于细菌人工染色体（BAC）的序列标签位点（STS）内容图谱，覆盖人类1号染色体1p35 - p36区域的35兆碱基。

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

作者信息

Chen Y Z, Hayashi Y, Wu J G, Takaoka E, Maekawa K, Watanabe N, Inazawa J, Hosoda F, Arai Y, Ohki M, Mizushima H, Morohashi A, Ohira M, Nakagawara A, Liu S Y, Hoshi M, Horii A, Soeda E

机构信息

Gene Bank, Tsukuba Institute, Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyadai, Tsukuba, 305-0074, Japan.

出版信息

Genomics. 2001 May 15;74(1):55-70. doi: 10.1006/geno.2001.6511.

DOI:10.1006/geno.2001.6511

PMID:11374902

Abstract

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.

摘要

我们设计了一种映射方法，用于在辐射杂种（RH）板上使用序列标签位点（STS）和聚合酶链式反应（PCR）对细菌人工染色体（BAC）克隆进行快速组装和排序。该方案包括两轮从有限数量的BAC中进行二维筛选，以使每个BAC都对应一个STS。在第一轮中，将STS组装到RH箱中，并根据来自已排列BAC克隆的384孔微量滴定板（MTP）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染色体1p35 - p36的35兆碱基（Mb）区域，并在11个重叠群中组装了1366个BAC，最长的约为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。