Carbon Nanotube Probes for Direct DNA Sequence Analysis

说明：（申请人提供） 人类基因组计划提供了大量的遗传信息 这将彻底改变对遗传性疾病的理解和诊断， 疾病特别是，希望单核苷酸的检测 基因编码区和调控区的多态性（SNP）将导致 更好地理解基因对癌症风险的贡献， 阐明影响治疗和预防的遗传因素，以及 强大新药的设计。实现这些目标需要 在包括检测或发现SNPs在内的多个领域做出了大量努力 和开发高通量的方法来表征和比较 基因组特定区域的核苷酸变异。此外，最近 观察表明，受试者的单倍型-特定等位基因 与每个染色体同源物相关-是SNP中的关键元件 映射，虽然目前用于确定单倍型的方法有显着 这些限制阻碍了它们在大规模遗传筛查中的应用。 为了解决这一重大技术差距，并提供关键信息， 为分子水平的癌症研究和医学，该项目将开发一个 一种新的技术，它是基于DNA的直接分子尺度成像 使用碳纳米管原子力显微镜探针，用于检测， 核苷酸变异的表征，特别是直接单倍型 保持战略定力在项目的R21阶段，我们将开发材料， 制造纳米管探针尖端所需的科学和化学， 可重复的检测能力，并整合生物学方法， DNA样品制备和沉积的化学和工程 适用于可重复分析。在随后的R33阶段， 项目，我们将大幅提高小说的吞吐量 （一）开发一个能够自动化的综合系统， 样品阵列的沉积、阵列成像和图像分析，（ii）系统和 显著增加样品检测通量的软件，以及（iii） 多探针阵列，用于高通量并行样品成像。

DESCRIPTION: (Provided by applicant) The Human Genome Project is providing massive amounts of genetic information that should revolutionize the understanding and diagnosis of inherited diseases. In particular, it is hoped that the detection of single nucleotide polymorphism (SNPs) in gene coding and regulatory regions will lead to a greater comprehension of the genetic contribution to risk for cancer, to the elucidation of the genetic factors that affect treatment and prevention, and to the design of powerful new drugs. Achieving these goals will require substantial effort in several areas, including detection or discovery of SNPs and development of high throughput methods for characterizing and comparing nucleotide variations in specific regions of the genome. In addition, recent observations suggest that the haplotype of a subject - the specific alleles associated with each chromosome homologue - is a critical element in SNP mapping, although current methods for determining haplotypes have significant limitations that have prevented their use in large-scale genetic screening. To address this substantial technology gap and provide critical information for molecular level cancer research and medicine, this project will develop a novel technology, which is based upon direct molecular scale imaging of DNA using carbon nanotube atomic force microscopy probes, for the detection and characterization of nucleotide variations, and in particular, direct haplotype determination. In the R21 phase of the project, we will develop the materials science and chemistry required to make nanotube probe tips that provide reproducible detection capability, and to integrate methods from biology, chemistry and engineering for preparation and deposition of DNA samples suitable for reproducible analysis. In the subsequent R33 phase of the project, we will be extend significantly the throughput of our novel technology by developing (i) an integrated system capable of automated deposition of sample arrays, array imaging and image analysis, (ii) system and software to increase significantly sample detection throughput, and (iii) multiprobe arrays for ultrahigh throughput parallel sample imaging.