Ordering of Chromosome Specific Overlapping Cosmid Clones

染色体特异性重叠粘粒克隆的排序

• 批准号: 9122035
• 负责人: William MacConnell
• 金额: $ 26万
• 依托单位: MacConnell Research
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 1994-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9122035&HistoricalAwards=false
• 关键词: Ordering; Chromosome; Specific; Overlapping; Cosmid

In the phase II work proposed research we will determine the overlapping relationships between-10,000 clones from a cosmid library derived from partially-digested, chromosome-sorted human DNA. This information will allow the creation of an ordered chromosome specific library. This library, and others to follow, will be indispensable tools for the human genome mapping and sequencing project. The phase II work will create an ordered cosmid library as well as kits, devices, and instruction manuals that can be used by other investigators to accomplish the same result on segments of DNA as large as entire human chromosomes. The phase II work involves cross-hybridization of the inserts from a large number of cosmid clones to each other independently in all combinations using the "pen-line" method developed in phase I work. The "pen-line" method allows discrete hybridization of all cloned DNAs separately and independently to all probes generated from the same set clones and, therefore, allows for simultaneous determination of the overlap between all clones in the set. The method is several fold faster than current repeatable protocol. In phase II work, we propose to use the method on an overlapping clone library derived from chromosome 20.

在第二阶段的工作提出的研究，我们将确定 一个粘粒中约10，000个克隆之间的重叠关系 来源于部分消化的染色体分选的人的文库 DNA. 这些信息将允许创建一个有序的 染色体特异性文库 这个图书馆，以及其他图书馆， 将成为人类基因组图谱绘制不可或缺的工具， 测序项目。 第二阶段的工作将建立一个有序的 粘粒库以及试剂盒、设备和说明书 其他研究人员也可以使用它来完成同样的任务 结果是整个人类染色体大小的DNA片段。 第二阶段的工作包括交叉杂交的插入从 大量的粘粒克隆相互独立， 使用第一阶段工作中开发的“笔线”方法进行组合。 “笔线”方法允许所有克隆的杂交。 DNA分别和独立于所有探针产生的 相同的克隆集，因此允许同时 确定集合中所有克隆之间的重叠。 的 该方法比当前的可重复协议快几倍。 在第二阶段的工作中，我们建议使用重叠的方法 来自20号染色体的克隆文库。