PHOTOCHEMICAL MODIFICATION OF HYBRIDIZATION MEMBRANES

杂交膜的光化学改性

• 批准号: 3498695
• 负责人: LISE W DURAN
• 金额: $ 5万
• 依托单位: SURMODICS, INC.
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-04-01 至 1992-12-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3498695
• 关键词: DNA; acrylamides; biomaterial development /preparation; cellulose; chemical binding; chemical group; chemical stability; chemical synthesis; copolymer; covalent bond; ionic bond; membrane; method development; nucleic acid hybridization; nucleic acid probes; photoactivation; photochemistry; polyvinyls

DESCRIPTION (Adapted from applicant's abstract):The transfer of nucleic acids to membrane filters has provided a number of applications for molecular hybridization including the analysis of genome structure and function and diagnosis of infectious diseases and genetic disorders. The sensitivity and reproducibility of the hybridization reaction depend on the properties of the membrane used.BSI has developed a proprietary photoimmobilization technology that allows covalent coupling of reagents onto numerous membrane materials. It is proposed to apply this technology to modify the surface of membranes used in nucleic acid hybridization assays. The Phase I objectives are to synthesize coating reagents which will permit the attachment of photoreactive groups to membrane surfaces that are capable of covalently bonding to DNA; demonstrate that positively charged groups used to ionically bind the DNA can be removed after DNA photoimmobilization is complete to reduce background problems; and define conditions for DNA transfer, and hybridization and stripping of probes on modified membranes. The long-range goal is to develop a membrane that can firmly bind DNA fragments of all sizes, provide maximum signal with minimal background, can be reprobed multiple times, and can be used in a number of formats under a wide range of conditions.

描述（改编自申请人摘要）：核酸转移 酸到膜过滤器提供了许多应用， 分子杂交，包括基因组结构分析， 传染病和遗传病的功能和诊断。 的 杂交反应的灵敏度和再现性取决于 BSI开发了一种专有的 允许试剂共价偶联的光固定化技术 在许多膜材料上。 建议应用这项技术 修饰核酸杂交中使用的膜的表面 分析。 第一阶段的目标是合成涂层试剂， 光反应性基团与膜表面的连接， 能够与DNA共价结合;证明带正电荷的 用于离子结合DNA的基团可以在DNA 完成光固定化以减少背景问题;并且 用于DNA转移的条件，以及探针的杂交和剥离。 改性膜 长期目标是开发一种膜， 牢固地结合所有大小的DNA片段，提供最大的信号， 背景，可以多次重新探测，并且可以用于许多 在广泛的条件下的格式。