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， 年 测序将通过正式连接我们的 位于俄克拉荷马州的一个实验室，与GESTEC合作进行制图 22号染色体位于费城，与约翰·萨尔斯顿博士和他的 英国欣克斯顿桑格中心的同事们。通过这些 合作将是共同协调努力， 整个染色体，以及努力提高效率， 序列数据收集、分析、注释和发布。 为了实现这一目标，我们现在建议1。扩展兆碱基序列 俄克拉荷马州大学基因组高级中心的能力 技术（ACGT）。 ACGT是建立在研究生/博士后 基于培训的基因组测序方法， 成功完成约100万个人类DNA碱基 在过去的一年里， 我们将生产至少2 Mb的成品 通过将我们现有的测序能力翻一番， ABI 373 A荧光测序仪4 ~ 8台。 然后在 未来两年每年增加4台新的测序仪器 随着预期的改进，将产生4 Mb的成品 序列在第2年，和5 Mb的完成序列在每年的3，4， 和5. 通过使用和改进我们已经建立的协议，我们 可以完成人类22号染色体的一半序列 在拟议的补助期内。 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 分析，从而提高我们的DNA测序效率。 3.快速向社区发布注释的DNA序列数据 继C. elegans范式概述在我们的合作 安排与桑格中心，欣克斯顿，英格兰。 利用现有的技术，我们可以清楚地记录下 根据我们自己的生产力，华盛顿大学（C。 elegans）和桑格中心（酵母，C. elegans和human）。 成本 最终序列的每个碱基略低于1美元， NCHGR向PI提供的资金总额，包括间接成本和按比例分摊 设备，并除以总数 在GenBank中已登录。 随着适度的改善， 技术和效率，我们可以现实地预计， 人类22号染色体的一半可以在计划的5年内完成测序 赠款期间的成本接近50美分，每个基地的最后序列。 在这期间，我们还将继续训练下一个 一代科学家的基本理论和方法需要， 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.