DNA SEQUENCING AND ANNOTATION OF HIGH RESOLUTION

描述：（根据申请人摘要改编）项目目标 2是将Project产生的基于重叠群的序列就绪图 1转换为详细的高分辨率注释序列图， DNA测序 与项目1并行运行，使用 为人类染色体11、21、20、10、13、14和15生成的Cosmopolymers图谱。 根据第18和第8条，该项目将以高度自动化的方式进行。 由项目1产生的序列就绪重叠群图谱，由10个 到20倍覆盖的粘粒末端，将受到高通量 直接从粘粒模板进行DNA测序，使用自动化 模板DNA制备、自动化反应组装和孵育， 使用常规荧光的自动序列测定 技术，并使用定制的 并行处理微型计算机 这一分析将导致 每个染色体约三分之一的一次通过序列， 长度为350至550 bp的序列片段，具有1至5 kb的缺口， 并将作为完成DNA测序的直接前奏。 这 将是确定和分析30%到50%的 染色体序列 利用现有的信息分析这些信息， 由计算核心开发的程序和附加软件 （1）建立基于物理的STS （2）检测至少100 kb的均匀间隔的位置; 足够的简单序列重复，以及它们向PCR的转化。 可检测的多态性标记，以将遗传图谱细化到至少0.1 基于物理位置的厘摩分辨率;以及（3）完整的 特别感兴趣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那些 人类遗传疾病的研究

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.