CHARACTERIZATION AND QUANTIFICATION OF IMMOBILIZED DNA

固定化 DNA 的表征和定量

• 批准号: 6481229
• 负责人: FEIMENG ZHOU
• 金额: $ 7.32万
• 依托单位: CALIFORNIA STATE UNIVERSITY LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6481229
• 关键词: DNA; atomic force microscopy; biosensor device; electrochemistry; mass spectrometry; membrane reconstitution /synthesis; metal complex; molecular assembly /self assembly; molecular film; nucleic acid probes; nucleic acid quantitation /detection; oligonucleotides; polynucleotides; scanning electron microscopy

In the proposed work, various schemes for fabricating well-ordered oligonucleotide and polynucleotide films onto gold surfaces will be explored. Emphasis will be placed on the formation of self-assembled monolayers (SAMs) containing DNA molecules or attaching DNA molecules onto preformed SAMs of alkanethiols. These films will be quantified and characterized using a variety of surface and analytical techniques. Five techniques will be employed in an effort to provide a detailed description about the heterogeneous DNA sensor surface: quartz crystal microbalance, atomic force microscope, scanning electrochemical microscope, voltammetry, and electrochemical inductively coupled plasma (ICP)-atomic emission spectrometry ICP-mass spectrometry. These techniques complement one another in terms of studies of DNA immobilization, hybridization intercalation, and film desorption. We intend to integrate the design of the DNA immobilization scheme, the characterization of the sensing surface, and the analytical performance of the resulting sensor of the resulting sensor. Our ultimate objective is to understand certain important factors governing DNA sensor performance (e.g., surface coverage of immobilized DNA, DNA molecular orientation, optimal concentrations of the probe and target for an analysis, and the development of an amenable technique for sensitive and selective detection of nucleic acids) at molecular levels.

在拟议的工作中，将探索在金表面上制造有序寡核苷酸和多核苷酸薄膜的各种方案。重点将放在包含 DNA 分子的自组装单层 (SAM) 的形成或将 DNA 分子附着到预先形成的烷硫醇 SAM 上。这些薄膜将使用各种表面和分析技术进行量化和表征。将采用五种技术来详细描述异质 DNA 传感器表面：石英晶体微天平、原子力显微镜、扫描电化学显微镜、伏安法和电化学感应耦合等离子体 (ICP)-原子发射光谱法、ICP-质谱法。这些技术在 DNA 固定、杂交嵌入和薄膜解吸研究方面相互补充。我们打算整合 DNA 固定方案的设计、传感表面的表征以及最终传感器的分析性能。我们的最终目标是了解控制 DNA 传感器性能的某些重要因素（例如，固定化 DNA 的表面覆盖、DNA 分子方向、用于分析的探针和靶标的最佳浓度，以及开发用于敏感和选择性检测的可靠技术）核酸）在分子水平上。