Single molecule DNA/RNA transport and Raman scattering readout in a coupled nanochannel/nanopore sequencing system.

耦合纳米通道/纳米孔测序系统中的单分子 DNA/RNA 传输和拉曼散射读数。

• 批准号: 10155991
• 负责人: Steven Brueck
• 金额: $ 34.92万
• 依托单位: ARMONICA TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-17 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10155991
• 关键词: 3-Dimensional; Alpha Particles; Area; Binding Proteins; Caliber; Chemicals; Coupled; DNA; DNA Binding; DNA Modification Process; DNA sequencing; Deposition; Detection; Deuterium; Development; Dimensions; Electromagnetic Fields; Electromagnetics; Engineering; Enzymes; Epigenetic Process; Genomics; Goals; Homo; Hot Spot; Individual; Isotopes; Label; Lasers; Measurement; Medical; Messenger RNA; Metals; Methylation; Modification; Nanostructures; Noise; Nucleotides; Oligonucleotides; Optics; Particle Size; Pattern; Phase; Process; Quantum Dots; RNA; RNA Transport; Resolution; Signal Transduction; Source; Streptavidin; Stretching; Structure; Surface; System; Technology; Testing; Time; Transcription Initiation Site; Variant; atomic layer deposition; base; colloidal nanoparticle; commercialization; density; ds-DNA; electric field; experimental study; improved; lithography; nanochannel; nanoparticle; nanopore; nanoscale; novel; operation; plasmonics; seal; self assembly; sequencing platform; simulation; single molecule; vapor; vibration

Single molecule DNA/RNA transport and Raman Scattering readout in a coupled nanochannel/nanopore sequencing system Armonica Technologies, Inc. is proposing to develop a novel, high-throughput, label-free, highly accurate, long-read DNA sequencing platform based on inexpensive nanoscale patterning and self- assembly. The platform consists of nanochannels (cross section dimensions of ~ 100 nm); tortuous (convoluted, 3D) nanopores formed by self-assembly of colloidal nanoparticles; nanoparticle barriers placed across the nanochannels; and a metal-insulator-metal (MIM) field enhancement structure atop the nanochannel roof. In operation, single- stranded- or double-stranded-DNA is partially stretched into a linear configuration in the nanochannels, is blocked at barriers incorporated into the channels and forced (by electric field) to translocate through the tortuous nanopores in the roof. The MIM structure on the roof locally enhances the electromagnetic fields of applied laser sources allowing surface enhanced coherent anti-Stokes Raman scattering (SECARS) detection of individual bases as they pass through the electromagnetic hot spots, thus providing single base sensitivity and spatial localization. The distinct Raman spectra of the individual bases allow label-free sequencing. Optical detection allows massively parallel operation since the only requirement is separation of the pores by more than an optical wavelength, which is easily accomplished in the fabrication. An important feature of the platform is that the porous roofs allow introduction of oligonucleotides, small proteins, and DNA-binding/DNA processing enzymes, permitting optional manipulation and modification of the DNA in the nanochannels. The goals of this Phase I project are: to optimize the MIM structure for single base sensitivity and spatial localization; to demonstrate single base sensitivity

耦合中的单分子 DNA/RNA 传输和拉曼散射读数 纳米通道/纳米孔测序系统 Armonica Technologies, Inc. 提议开发一种新型、高通量、无标签、高度 准确、长读长的 DNA 测序平台，基于廉价的纳米级图案和自 集会。该平台由纳米通道组成（横截面尺寸约为 100 nm）；曲折 （卷积，3D）由胶体纳米粒子自组装形成的纳米孔；纳米粒子屏障 放置在纳米通道上；以及顶部的金属-绝缘体-金属（MIM）场增强结构 纳米通道屋顶。在操作中，单链或双链 DNA 被部分拉伸成 纳米通道中的线性构型被纳入通道中的屏障阻挡并被迫 （通过电场）通过屋顶上曲折的纳米孔进行移位。屋顶MIM结构 局部增强所应用激光源的电磁场，从而实现表面增强相干 当单个碱基通过时，进行反斯托克斯拉曼散射 (SECARS) 检测 电磁热点，从而提供单碱基灵敏度和空间定位。独特的 各个碱基的拉曼光谱允许无标记测序。光学检测允许大规模 并行操作，因为唯一的要求是通过多于光学的方法来分离孔 波长，这在制造中很容易实现。该平台的一个重要特点是 多孔屋顶允许引入寡核苷酸、小蛋白质和 DNA 结合/DNA 加工酶，允许对纳米通道中的 DNA 进行选择性操作和修饰。 该第一阶段项目的目标是：优化MIM结构以实现单碱基敏感性和空间 本土化;证明单碱基敏感性