HIGH FIDELITY SURFACES AND DNA ARRAYS

描述（研究者摘要）：本提案的主要目标是 开发高质量的表面和核酸阵列， 包括基因表达，遗传变异， 蛋白质：核酸的相互作用，光学绘图。拟议工作 福尔斯分为两个相互关联的领域：各种表面的开发 为特定应用而设计和优化;在此基础上， 研究，以提高对化学物质的理解和控制， 自己浮出水面。待开发的表面分为三大类 类别：均匀（未阵列）表面、在正方形网格中阵列的表面 图案和纳米结构表面。表面将采用以下材料进行处理： 由应用确定的各种化学官能度。这些 官能团将包括羟基（光导向DNA的底物 阵列制造）;氨基（光学映射的底物，非共价 PCR片段沉积或活化寡核苷酸的共价连接）; 硫醇基团（用于共价连接活化的 寡核苷酸）;和硫代反应性或氨基反应性基团（对于共价键）。 寡核苷酸或PCR扩增子的连接）。两个气相沉积的金 载玻片上的膜和原子级平坦的硅晶片将被用作 印刷受体. DNA分子将通过直接化学偶联 或者通过静电相互作用到气相沉积的金薄膜上， 用致密的自组装单分子层进行化学修饰， 用各种官能团封端的烷硫醇。附接到 原子级平坦的氢封端硅表面将通过 官能化脂肪族烯烃形成Si-C的UV介导反应 键，也适用于进一步修饰或直接DNA连接。的 表面将被要求满足严格的性能标准，包括 物理和化学稳定性，热稳定性， 杂交和酶促修饰， 表面结合DNA和制备的再现性。

DESCRIPTION (Investigator's Abstract): The principal goal of this proposal is the development of high quality surfaces and nucleic acid arrays for applications including the analysis of gene expression, of genetic variations, of protein: nuclei acid interactions, and optical mapping. The proposed work falls into two interrelated areas: the development of a variety of surfaces designed and optimized for particular applications; and underlying this, basic research to develop improved understanding and control of the chemistry of the surfaces themselves. The surfaces to be developed fall into three major categories: Uniform (unpatterned) surfaces, surfaces patterned in a square grid pattern, and nanostructured surfaces. The surfaces will be prepared with various chemical functionalities as determined by the application. These functionalities will include hydroxyl groups (substrates for light-directed DNA array fabrication); amino groups (substrates for optical mapping, noncovalent PCR fragment deposition, or covalent attachment of activated oligonucleotides); thiol groups (substrates for covalent attachment of activated oligonucleotides); and thio-reactive or amino-reactive groups (for covalent attachment of oligonucleotides or PCR amplicons). Both vapor-deposited gold films on glass slides, and atomically flat silicon wafers will be employed as substrates. DNA molecules will be attached either by direct chemical coupling or by electrostatic interactions onto vapor-deposited gold thin films that have been chemically modified with densely packed self-assembled monolayers of alkanethiols that are terminated with various functional groups. Attachment to atomically flat hydrogen-terminated silicon surfaces will be accomplished by the UV-mediated reaction of functionalized aliphatic alkenes to form Si-C bonds, also suitable for further modification or direct DNA attachment. The surfaces will be required to meet stringent performance criteria, including physical and chemical stability, thermal stability, amenability to hybridization and enzymatic modification, control of the density of surface-bound DNAs, and reproducibility of preparation.