PHYSICAL MAPPING COMPONENT

人类基因组计划现在的重点是获得完整的序列 我们的基因组。只有掌握了这些信息， 构成人类蓝图的所有基因和调控元件。 任何测序项目的开始都需要合适的模板。为 大规模的测序工作，显然是一个不平凡的事情。 特别是，人类基因组的物理图谱不包括 容易测序的克隆。这些基于YAC的STS内容映射和 辐射混合地图没有分辨率和标记密度， 使BAC和/或PAC叠连群的组装成为可能。在没有 序列准备好的地图，必须以一种能够维持 测序工作与一个不间断的流动，以及表征 克隆需要足够的重叠群深度以确保重叠群的完整性。 选择克隆以代表最小平铺路径。测序 如果选择删除或重排的克隆， the representation代表of the genome基因. 这个项目的具体目标是组装一个序列准备好的地图， 人类18号染色体，以支持大规模测序工作， 人类基因组中8500万碱基的片段。这将是 分两个阶段完成。第一阶段将侧重于标记密集的 区域的染色体，以便快速生成BAC的选择 克隆来启动测序工作一个完成了，第二个 然后，阶段将致力于构建序列就绪的 整个染色体缩小差距和保持连续性将是首要目标 使得可以选择有效的平铺路径以减少 大量的冗余序列。BAC克隆最初将通过以下方法分离： 将公众可获得的作图标记与高密度过滤器杂交 克隆人阵列为了关闭，将使用 从末端游走序列开发的探针和来自 选择的YAC克隆。所有克隆将使用完整的指纹 限制性内切酶酶切和琼脂糖凝胶电泳。康蒂格斯会 使用可识别克隆重叠的FPC软件包进行组装 通过比较限制模式的相似性。

The Human Genome Project is now focused on obtaining the complete sequence of our genome. It is only with this information in hand can we identify all the genes and regulatory elements that make up the human blueprint. The start of any sequencing project requires a suitable template. For large-scale sequencing efforts, the obvious point is a non-trivial matter. In particular, the physical maps of the human genome are not comprised of clones which are readily sequenced. These YAC-based STS content maps and radiation hybrid maps do not have the resolution and marker density to enable ready assembly of BAC and/or PAC contigs. In the absence of a sequence-ready map, one must be constructed at a rate that will sustain the sequencing effort with an uninterrupted flow of well characterized clones. Sufficient contig depth is needed to ensure the integrity of the clones selected to represented the minimal tiling path. Sequencing accuracy is a moot point if deleted or rearranged clones are selected as the representation of the genome. The specific aim of this project is to assemble a sequence-ready map of human chromosome 18 in support of a large-scale sequencing effort directed at sequencing this 85 megabase segment of the human genome. This will be accomplished in two phases. The initial phase will focus on a marker-dense region of the chromosome in order to generate quickly a selection of BAC clones to jump-start the sequencing effort. One accomplished, the second phase will then be directed at constructing a sequence-ready map of the entire chromosome. Gap closure and continuity will be a prime objective such that an efficient tiling path can be selected in order to reduce the amount of redundant sequencing. BAC clones will be initially isolated by hybridization of publicly available mapped markers to high-density filters of arrayed clones. For closure, additional clones will be isolated using probes developed from end-walk sequences and random reads derived from selected YAC clones. All clones will be finger-printed using complete restriction enzyme digests and agarose gel electrophoresis. Contigs will be assembled using the FPC software package that identifies clone overlap by comparing restriction patter similarities.