DNA SEQUENCING, TESTING THE OSS APPROACH AND RELATED TECHNOLOGY

基于YAC的7号染色体重叠克隆/STS/遗传连锁图谱将 是被建造的。将从200到200的序列中开发出一对引物 1000个碱基的染色体特异DNA克隆，通过流动分选获得 染色体，并将用于PCR反应以筛选人的文库 同源克隆的YAC中的DNA。重叠的YAC将用于获得 平均长度至少为2Mb的重叠群根据其探针含量排列。 将做出辅助努力来识别YAC并从以下位置构建重叠群 7号染色体上当前感兴趣区域中的现有探针(例如， 克隆的基因)。关闭图谱，并将其与Genetic和 细胞遗传学数据，将首先基于从 YAC重叠群末端，以及这些区域的多态鉴定 关于DNA的。末端克隆的多态将使用以下方法进行遗传作图 子代的染色体从一组初级的 参考家系，这应该允许快速比对 相应的重叠群和基因座在遗传连锁中的放置 地图。地图的闭合将通过映射衍生的多态来实现 来自端粒YAC和来自着丝粒周围区域的克隆DNA，并通过 跳跃克隆、原位杂交和脉冲场凝胶的应用 映射。结果将是一张地图，在其中，细胞遗传学分配， 遗传座位的连锁信息，以及重叠克隆的集合 完整描述了7号染色体的基因被整合在一起。

A YAC-based overlapping clone/STS/genetic linkage map of chromosome 7 will be constructed. Primer pairs will be developed from the sequence of 200 to 1000 bp clones of chromosome specific DNA, obtained from flow-sorted chromosomes, and will be used in PCR reactions to screen a library of human DNA in YACs for cognate clones. Overlapping YACs will be used to obtain contigs of an average length at least 2 Mb aligned by their probe content. Subsidiary efforts will be made to identify YACs and build contigs from existing probes in regions of current interest on chromosome 7 (e.g., cloned genes). Closure of the map, and its merger with genetic and cytogenetic data, will be based first on the generation of clones from the ends of YAC contigs, and identification of polymorphisms in these regions of DNA. End-clone polymorphisms will be genetically mapped using crossovers in the chromosomes of the offspring from a primary set of reference pedigrees, which should allow rapid alignment of the corresponding contigs and placement of the loci within the genetic linkage map. Closure of the map will be attained by mapping polymorphisms derived from telomeric YACs and cloned DNAs from the pericentromeric region, and by the use of jumping clones, in situ hybridization and pulsed-field gel mapping. The result will be a map in which cytogenetic assignments, linkage information for genetic loci, and the set of overlapping clones that fully describe chromosome 7 are integrated.