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

单分子DNA/RNA输运和拉曼散射读出 纳米通道/纳米孔测序系统 Armonica Technologies，Inc.提出开发一种新的，高通量，无标记，高度 基于廉价的纳米级图案化和自组装的精确、长读段DNA测序平台， 组装件.该平台由纳米通道（横截面尺寸约100 nm）组成;曲折 通过胶体纳米颗粒的自组装形成的（回旋的，3D）纳米孔;纳米颗粒屏障 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被部分拉伸成单链DNA。 纳米通道中的线性构型在结合到通道中的屏障处被阻断， (by电场）以移位通过顶部中的曲折纳米孔。屋顶上的MIM结构 局部增强所施加的激光源的电磁场， 反斯托克斯拉曼散射（SECARS）检测单个碱基，因为它们通过 电磁热点，从而提供单碱基灵敏度和空间定位。鲜明的 单个碱基的拉曼光谱允许无标记测序。光学检测可以大规模地 平行操作，因为唯一的要求是通过超过光学孔径的孔径来分离孔， 波长，这在制造中容易实现。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，允许任选地操纵和修饰纳米通道中的DNA。 第一阶段项目的目标是：优化MIM结构的单碱基灵敏度和空间分辨率。 定位;证明单碱基灵敏度