SEQUENCING HUMAN CHROMOSOME 22, CENTROMERE TO NF2

本课题研究的主要目标是系统地 在接下来的5年内对人类22号染色体的大约一半进行测序 好几年了。测序将通过正式链接我们的 总部设在俄克拉何马州的实验室与GESTEC合作进行测绘 22号染色体总部设在费城，与约翰·苏斯顿博士和他的 英国辛克斯顿桑格中心的同事们。通过这些 合作将共同协调努力，以排序 整个染色体，以及提高效率的努力 序列数据收集、分析、注释和发布。 为了实现这一目标，我们现在提出1.扩展兆库测序 俄克拉荷马大学高级基因组中心的能力 技术(ACGT)。ACGT是建立在研究生/博士后基础上的 基于培训的基因组测序方法已经导致了 成功地完成了大约100万个人类DNA碱基 在过去一年内的序列。我们将生产至少2Mb的成品 通过将我们现有的测序能力加倍，在第一年进行测序 4~8台ABI 373A荧光测序仪。然后，使用 在未来两年每年增加4台新的测序仪器 加上预期的改进，将生产4Mb的成品 第二年的序列，以及第三、四、四年的每年5Mb的完成序列， 5.通过采用和完善我们已经建立的协议，我们 可以完成人类22号染色体序列的大约一半 在拟议的授权期内。 2.继续发展、改进和实施自动化程序 用于DNA分离、DNA序列分析、数据采集和数据 分析，从而提高我们的DNA测序效率。 3.向社区快速发布带注释的DNA序列数据 遵循我们的合作项目中概述的线虫范例 与英国辛克斯顿桑格中心的安排。 利用现有的技术，我们可以清楚地记录这些预测 根据我们自己的生产率，华盛顿大学(C. 和桑格中心(酵母、线虫和人类)。成本 最终序列的每个碱基略低于1美元，计算方法为 NCHGR对PI的总资金，包括间接成本和按比例分配 整个授权期内的设备数量，并除以总数 O~碱基存放在GenBank中。随着以下方面的适度改进 技术和效率，我们可以现实地预计大约 人类22号染色体的一半可以在拟议的5年内测序 授权期，最终序列的每个碱基成本接近50美分。 在这段时间里，我们还将继续培训下一位 一代科学家在基础理论和方法方面需要 发展新的测序和数据分析方法。

The major goal of the research in this project is to systematically sequence approximately half of human chromosome 22 within the next 5 years. The sequencing will be accomplished by officially linking our laboratory based in Oklahoma in a partnership with the GESTEC for Mapping Chromosome 22 based in Philadelphia and with Dr. John Sulston and his colleagues at the Sanger Center, Hinxton, England. Through these collaborations will be jointly coordinating efforts to sequence the entire chromosome, as well as efforts to improve the efficiency of sequence data collection, analysis, annotation, and release. To achieve this goal, we now propose to 1. expand the megabase sequencing capabilities at the University of Oklahoma Advanced Center for Genome Technology (ACGT). ACGT is built upon the graduate student/postdoctoral training-based approach to genomic sequencing that has led to successfully completing approximately 1 million bases of human DNA sequence within the past year. We will produce at least 2 Mb of finished sequence in year 1 by doubling our existing sequencing capabilities, from 4 to 8 ABI 373A fluorescent sequencing instruments. Then, with the addition of 4 new sequencing instruments in each of the next two years and with the anticipated improvements, will produce 4 Mb of finished sequence in year 2, and 5 Mb of finished sequence in each of years 3, 4, and 5. By employing and refining our already established protocols, we can complete approximately one-half the sequence of human chromosome 22 within the proposed grant period. 2. To continue to develop, improve and implement the automated procedures for DNA isolation, DNA sequence analysis, data acquisition, and data analysis, thereby increasing our DNA sequencing efficiency. 3. To rapidly release annotated DNA sequence data to the community following the C. elegans paradigm as outlined in our collaborative arrangement with the Sanger Center, Hinxton, England. With the existing technology, we can clearly document the predictions based on our own productivity, that of the Washington University (C. elegans), and the Sanger Center (yeast, C. elegans, and human). The cost per base of final sequence is slightly under $1 as calculated by taking the total NCHGR funding to the PI, including indirect cost and pro-rating equipment over the total grant period, and dividing by the total number o~ bases deposited in GenBank. With the modest improvements in technology and efficiency, we can realistically expect that approximately half of human chromosome 22 can be sequenced within the 5 year proposed grant period at a cost approaching 50 cents per base of final sequence. Throughout this period, we also will continue to train the next generation of scientists in the basic theories and methods needed to evolve new approaches to sequencing and to data analysis.