DNA SEQUENCING AND ANNOTATION OF HIGH RESOLUTION

高分辨率 DNA 测序和注释

• 批准号: 6109076
• 负责人: GLEN A EVANS
• 金额: $ 126.32万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6109076
• 关键词: chromosome 21; chromosomes; computer assisted sequence analysis; genetic mapping; human genetic material tag; nucleic acid sequence; parallel processing; plasmids; sequence tagged sites

Description: (Adapted from the applicant's abstract) The goal of Project 2 is to convert the contig-based sequence-ready maps produced by Project 1 into detailed high-resolution annotated sequence-based maps by selected DNA sequencing. Operating in parallel with Project 1, using the set of cosmids maps generated for human chromosomes 11, 21, 20, 10, 13, 14 and 18 and 8, this project will be carried out in a highly automated manner. The sequence-ready contig maps produced by Project 1, consisting of 10 to 20X coverage of the cosmid ends, will be subjected to high-throughput DNA sequencing directly from the cosmid templates, using automated template DNA preparation, automated reaction assembly and incubation, automated sequence determination using conventional fluorescence technology, and immediate on-line sequence analysis using custom designed parallel processing microcomputers. This analysis will result in the one-pass sequence of about one-third of each chromosome in ordered sequence fragments of 350 to 550 bp in length, with gaps of 1 to 5 kb, and will serve as an immediate prelude to complete DNA sequencing. This will be the determination and analysis of 30 to 50 percent of the entire chromosome sequence. The analysis of this information using available programs and additional software to be developed by the computation core should result in: (1) The construction of STSs based upon physical position with an even spacing of at least 100 kb; (2) The detection of sufficient simple sequence repeats, and their conversion to PCR- detectable polymorphic markers, to refine the genetic map to at least 0.1 centimorgan resolution based upon physical position; and (3) The complete genomic or cDNA sequence of putative genes of exceptional interest, such as those encoding unique proteins or likely to represent candidate genes for human genetic diseases.

描述：（改编自申请人的摘要）项目的目标 2 是转换 Project 生成的基于重叠群的序列就绪图谱 1 进入详细的高分辨率注释的基于序列的图谱 DNA 测序。 与项目 1 并行运行，使用一组 为人类染色体 11、21、20、10、13、14 和 生成粘粒图谱 18和8日，该项目将以高度自动化的方式进行。 项目 1 生成的序列就绪重叠群图谱，包含 10 个 粘粒末端覆盖率达到 20 倍，将进行高通量处理 使用自动化技术直接从粘粒模板进行 DNA 测序 模板 DNA 制备、自动化反应组装和孵育、 使用传统荧光进行自动序列测定 技术，以及使用定制设计的即时在线序列分析 并行处理微型计算机。 该分析将导致 每条染色体约三分之一的一次性序列 长度为 350 至 550 bp 的序列片段，间隙为 1 至 5 kb， 并将作为完成 DNA 测序的前奏。 这 将确定和分析整个数据的 30% 至 50% 染色体序列。 使用现有信息对这些信息进行分析 由计算核心开发的程序和附加软件 应该导致：（1）基于物理的STS的构建 位置均匀，间距至少为 100 kb； (2) 检测 足够的简单序列重复，并将它们转换为PCR- 可检测的多态性标记，将遗传图谱细化至至少 0.1 基于物理位置的厘摩分辨率； (3) 完整的 具有特殊意义的推定基因的基因组或 cDNA 序列，例如 作为编码独特蛋白质或可能代表候选基因的蛋白质 对于人类遗传疾病。