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。提议开发一种新颖，高通量，无标签，高度 基于廉价的纳米级图案和自我的准确，长阅读的DNA测序平台 集会。该平台由纳米通道组成（约100 nm的横截面尺寸）；曲折 （卷曲，3D）由胶体纳米颗粒自组装形成的纳米孔；纳米颗粒屏障 放置在纳米通道上；以及在 纳米通道屋顶。在运行中，单链或双链DNA部分伸展到 纳米通道中的线性构型在融合到通道并强迫的障碍物中被阻塞 （通过电场）通过屋顶上的曲折纳米孔转移。屋顶上的MIM结构 局部增强了施加激光源的电磁场，使表面增强了 反stokes拉曼散射（secars）在单个基地通过时检测 电磁热点，从而提供了单个碱基灵敏度和空间定位。独特的 单个碱基的拉曼光谱允许无标签的测序。光学检测允许大规模检测 并行操作，因为唯一的要求是将毛孔分离不超过光学 波长，在制造中很容易完成。该平台的一个重要特征是 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DNA的可选操作和修饰。 该阶段I项目的目标是：优化单个基础灵敏度和空间的MIM结构 本土化;展示单个基础灵敏度