High Integrity and High Yield DNA Extraction Using a Nanostructured Surface

使用纳米结构表面进行高完整性和高产量 DNA 提取

基本信息

  • 批准号:
    8646703
  • 负责人:
  • 金额:
    $ 17.51万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2014
  • 资助国家:
    美国
  • 起止时间:
    2014-04-05 至 2015-09-24
  • 项目状态:
    已结题

项目摘要

High Integrity and High Yield DNA Extraction Using a Nanostructured Surface DNA/RNA extraction is a critical first step that underpins nearly all of molecular biology. Phenol/chloroform precipitation remains the gold standard when the highest DNA integrity, yield, and purity are needed. However, the significant manual skill and dexterity required makes this method labor intensive, low throughput, and nearly impossible to automate. While commercial kits and automated instruments using silica columns and beads are capable of high yield, these methods generally result in lower DNA integrity (i.e. more sheared, smaller fragments) and lower purity than well performed phenol/chloroform extractions. Existing silica technologies use porous gels, tightly packed columns, and microparticles to increase surface area, leading to reduced DNA integrity due to high shear forces imparted by flow and mixing. We propose a novel silica coated nanomembrane that uses a hierarchical structure of microscale ridges covered by nanoscale chips to create a flat, non-porous substrate with high surface area. This nanomembrane is easily fabricated using an inexpensive thermoplastic material (i.e. pennies per foot) and is capable of binding >20 mg of DNA per cm2. The nanoscale surface topography enables high surface area silica based DNA extraction while eliminating flow and particle based shear forces to facilitate high yield and high purity DNA recovery with exceptional DNA integrity. DNA is bound by simply allowing the sample to contact the membrane surface rather than flow through it. In Aim 1, we will fabricate a thermoplastic nanomembrane with a high density of nanoscale surface topography to create an inexpensive silica substrate with high surface area for DNA extraction. In Aim 2, we will develop an optimized protocol and buffer set for DNA extraction from human cell lines to obtain high DNA integrity, high extraction yield, and high purity. In Aim 3, we will perform DNA extractions using the nanomembrane, commercial spin-columns, magnetic beads, and phenol/chloroform to benchmark the optimized nanomembrane against existing methods. Through this project, we will develop a DNA extraction nanomembrane that combines the convenience of spin columns with the performance of phenol/chloroform.
利用纳米结构表面提取高完整性和高产率的DNA

项目成果

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Kelvin Liu其他文献

Kelvin Liu的其他文献

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{{ truncateString('Kelvin Liu', 18)}}的其他基金

Rapid Method to Enhance and Shape Long-Read Sequencing Read Length Distributions
增强和塑造长读长测序读长分布的快速方法
  • 批准号:
    10080760
  • 财政年份:
    2020
  • 资助金额:
    $ 17.51万
  • 项目类别:
Rapid Method to Enhance and Shape Long-Read Sequencing Read Length Distributions
增强和塑造长读长测序读长分布的快速方法
  • 批准号:
    10228764
  • 财政年份:
    2020
  • 资助金额:
    $ 17.51万
  • 项目类别:
Nanostructured Magnetic Disks for Wide Range Size Selection in NGS Library Preparation and PCR Purification
用于 NGS 文库制备和 PCR 纯化中宽范围尺寸选择的纳米结构磁盘
  • 批准号:
    9892018
  • 财政年份:
    2017
  • 资助金额:
    $ 17.51万
  • 项目类别:
Nanostructured Magnetic Disks for Wide Range Size Selection in NGS Library Preparation and PCR Purification
用于 NGS 文库制备和 PCR 纯化中宽范围尺寸选择的纳米结构磁盘
  • 批准号:
    10212466
  • 财政年份:
    2017
  • 资助金额:
    $ 17.51万
  • 项目类别:
Nanobind Hierarchical Silica Lamella for High Molecular Weight DNA Extraction
用于高分子量 DNA 提取的 Nanobind 分层二氧化硅片层
  • 批准号:
    9150684
  • 财政年份:
    2015
  • 资助金额:
    $ 17.51万
  • 项目类别:
Nanobind FFPE DNA/RNA Extraction
Nanobind FFPE DNA/RNA 提取
  • 批准号:
    8755436
  • 财政年份:
    2014
  • 资助金额:
    $ 17.51万
  • 项目类别:
High Quality, High Integrity Nucleic Acid Extraction from FFPE Tissues
从 FFPE 组织中提取高质量、高完整性的核酸
  • 批准号:
    9393293
  • 财政年份:
    2014
  • 资助金额:
    $ 17.51万
  • 项目类别:
Ligo-miR - A Multiplex Assay Platform for Targeted miRNA Profiling
Ligo-miR - 用于靶向 miRNA 分析的多重检测平台
  • 批准号:
    8904533
  • 财政年份:
    2012
  • 资助金额:
    $ 17.51万
  • 项目类别:
Ligo-miR - A Multiplexed Single Molecule Ligation Assay for miRNA Profiling
Ligo-miR - 用于 miRNA 分析的多重单分子连接测定
  • 批准号:
    8550117
  • 财政年份:
    2012
  • 资助金额:
    $ 17.51万
  • 项目类别:
PicoSep - A Microfluidic Single Molecule Free Solution Hydrodynamic Separation Pl
PicoSep - 微流控单分子自由溶液流体动力分离 Pl
  • 批准号:
    8546244
  • 财政年份:
    2012
  • 资助金额:
    $ 17.51万
  • 项目类别:

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