MEGABASE SEQUENCING ON THE HUMAN X CHROMOSOME

来自人类X染色体的3000万碱基DNA序列将是 在三年内以不到35美分的成本完成 基地。数据将是高质量的，每个错误不到一个 一万个核苷酸。Xq28和Xp22的高分辨率余弦图 区域将由自动限制性内切酶消化产生 映射。选定的宇宙星将通过平衡的随机和 基于亚克隆的双端测序的定向策略， 将最大限度地减少冗余和对昂贵的寡核苷酸的依赖 遍历，避免重叠中过多的重复排序 地区。宇宙地图中的间隙和歧义将通过以下方式解决 分离可选的基因组克隆，并通过协调正在进行的部分 用地图进行测序。新技术促进大规模生产 将开发测序技术。高质量的简化施工 猎枪库和快速DNA模板准备计划将允许 前端步骤的部分自动化。新型荧光染料标记 寡核苷酸引物将使序列反应流线化，并 由此产生的信号增强将简化DNA碱基调用。这个 信息学基础设施，用于协调来自 粘粒作图、DNA测序、数据组装、验证、注解和 将简化数据库提交，以实现更高的吞吐量。在……里面 除了DNA序列，该项目还将为 扩展的程序，可以对整个人类进行及时的排序 基因组。

Thirty megabases of DNA sequence from the human X chromosome will be generated over three years at a cost of less than 35 cents per finished base. The data will be of high quality, with less than one error per 10,000 nucleotides. High resolution cosmid maps from the Xq28 and Xp22 regions will be generated by automated restriction endonuclease digestion mapping. Selected cosmids will be analyzed by a balanced random and directed strategy based upon double ended sequencing of subclones that will minimize redundancy and dependence on expensive oligonucleotide walks, and avoid excessive redundant sequencing in the overlapping regions. Gaps and ambiguities in the cosmid maps will be solved by isolating alternative genomic clones, and by coordinating ongoing partial sequencing with the mapping. New technologies to improve large scale sequencing will be developed. Simplified construction of high quality shotgun libraries and a rapid DNA template preparation scheme will allow partial automation of front end steps. Novel fluorescent dye labeled oligonucleotide primers will enable streamlined sequence reactions and the resulting signal enhancements will simplify DNA base calling. The informatics infrastructure for the coordination of all steps from the cosmid mapping, DNA sequencing, data assembly, validation, annotation and database submission will be streamlined to allow higher throughput. In addition to the DNA sequence, this project will generate a model for an expanded program that can allow timely sequencing of the whole human genome.