CHARACTERIZATION AND QUANTIFICATION OF IMMOBILIZED DNA

固定化 DNA 的表征和定量

• 批准号: 6344122
• 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: 1978
• 资助国家: 美国
• 起止时间: 1978-06-14 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6344122
• 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.

在提议的工作中，将探索在金表面上制造有序的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分子连接到预制的烷硫醇sam上。这些薄膜将被量化和表征使用各种表面和分析技术。本文将采用石英晶体微天平、原子力显微镜、扫描电化学显微镜、伏安法和电化学电感耦合等离子体-原子发射光谱法ICP-质谱法等五种技术对异质DNA传感器表面进行详细描述。这些技术在DNA固定、杂交插层和膜解吸方面的研究是互补的。我们打算整合DNA固定方案的设计，传感表面的表征，以及所得传感器的所得传感器的分析性能。我们的最终目标是在分子水平上了解控制DNA传感器性能的某些重要因素（例如，固定DNA的表面覆盖率，DNA分子取向，用于分析的探针和靶标的最佳浓度，以及用于敏感和选择性检测核酸的适用技术的发展）。