INTEGRATED MAP FOR CHROMOSOME 12

12 号染色体综合图谱

• 批准号: 2541515
• 负责人: Raju S. Kucherlapati
• 金额: $ 127.72万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2541515
• 关键词: chromosomes; genetic library; genetic mapping; genetic markers; human genetic material tag; molecular cloning; nucleic acid sequence; polymerase chain reaction; sequence tagged sites

DESCRIPTION (Applicant's Abstract): The goal of this project is to contribute to the determination of the accurate sequence of human chromosome 12. Towards this goal, we entered into a collaboration with the Baylor College of Medicine (BCM) Genome Center. We propose to generate a sequence ready map of chromosome 12 in the form of P1 artificial chromosome (PAC) and bacterial artificial chromosome (BAC) maps. During the first year of the proposed grant, we propose to construct a map for 20 Mbp of chromosome 12, implement a bioinformatics strategy and put in place a number of high throughput systems to reach a production level mapping. During years two and three, we propose to construct maps for 100 Mbp of chromosome 12 and provide them to the BCM Genome Center. Based on this throughput, the sequence ready maps of chromosome 12 will be completed by December 31, 2000 and the sequence will be completed shortly thereafter. The general approach to construct the maps is as follows. We will use the 100 Kbp resolution physical map of human chromosome 12 in the form of a YAC contig map that we constructed as the framework map. PCR products from ten to twelve consecutive non-polymorphic markers will be used to screen a 20X genomic PAC/BAC library. False positive will be eliminated by a PCR based screening assay and the clones will be assembled into contigs by STS-content. Marker density will be increased by sequencing ends of clones by a direct sequencing method we developed and developing new STSs. Gaps will be filled by screening libraries with end clones. Clones that can constitute minimal tiling paths will be identified by fingerprinting methods. All of the data and the minimal tiling path of clones will be electronically transmitted to the BCM Genome Center and made publicly available through a web site. A team of six mappers assisted by an Informatics team will produce 20 Mbp of maps during the first year. We will implement automated methods for several steps in the procedure and, during the second and third years, we propose to increase the throughput by 2X with only the addition of two FTEs to the overall team. During years 2 and 3, the cost of producing the maps is estimated to be no more than $.025/nucleotide.

描述（申请人摘要）：本项目的目标是 有助于人类染色体的精确测序 12. 为了实现这一目标，我们与贝勒大学合作， 医学院基因组中心（College of Medicine Genome Center） 我们建议生成一个序列 P1人工染色体（PAC）形式的12号染色体的现成图谱， 细菌人工染色体（BAC）图谱。 头一年期间 建议赠款，我们建议构建一个20 Mbp的染色体12的地图， 实施生物信息学战略，并制定若干高级战略， 通过系统来达到生产水平映射。 在两年中， 第三，我们建议构建12号染色体100 Mbp的图谱， 把它们提供给基因组中心。 基于此吞吐量， 2000年12月31日将完成12号染色体的测序图 并且该序列将在此后不久完成。 的一般方法 构建地图的方法如下。 我们将使用100 Kbp分辨率 人类12号染色体的YAC重叠群图形式的物理图， 作为框架图。 10 - 12个PCR产物 将使用连续的非多态性标记来筛选20 X基因组 PAC/BAC库。 假阳性将通过基于PCR的筛查消除 测定，并将克隆通过STS含量组装成重叠群。 标记 通过直接测序法对克隆的末端进行测序， 我们开发的测序方法和开发新的STS。 空白将被填补 用末端克隆筛选文库。 可以构成最低限度的克隆 将通过指纹方法来识别平铺路径。 的所有数据 克隆的最小平铺路径将被电子传输到 BCM基因组中心并通过网站公开提供。 一 一个由六名绘图人员组成的小组将在信息学小组的协助下， 第一年的地图。 我们将实现自动化的方法， 在第二年和第三年，我们建议 仅增加两名全职员工， 整体团队。 在第二年和第三年，制作地图的费用为 估计不超过0.025美元/核苷酸。