AUTOMATED HIGH SPEED DNA SEPARATIONS

自动化高速 DNA 分离

• 批准号: 2889709
• 负责人: ANDREW G EWING
• 金额: $ 18.74万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2889709
• 关键词: DNA; biomedical automation; biomedical equipment development; electrochemistry; genotype; method development; nucleic acid sequence; physical separation

DESCRIPTION: Development of high throughput DNA sampling, separation, and detection strategies are described for use in sequencing and genotyping. New technology is proposed for massively parallel, low-volume DNA separations. This will be accomplished by combining capillary sample introduction parallel separations in microfabricated chips, and electrochemical array detection to provide a single integrated system for separation and detection. Ultimately, this will be connected via a sampling capillary to an array of nanovials for introduction of large numbers of samples. The proposal is divided into four major parts which include 1) optimization of separations of DNA fragments in ultrathin channels with sieving buffers, 2) development of an improved interface to allow efficient sample transfer across the capillary-channel interface, 3) development of electrochemical array detection using Ru(bpy)32+ as an electrophore for dsDNA and ssDNA, and 4) application of the capillary transfer method and electrochemical array detection to DNA separations on chips. This will provide the means to rapidly place multiple samples in a very large number of channels (1000 parallel channels is feasible) on a single chip. The proposed technology promises to increase sample throughput by 10- to 50-fold and reduce sample size by 2 to 3 orders of magnitude relative to currently available systems.

描述：开发高通量DNA采样、分离和纯化技术。 描述了用于测序和基因分型的检测策略。 提出了大规模并行、低容量DNA的新技术 分居 这将通过将毛细管样品 在微加工芯片中引入平行分离，以及 电化学阵列检测提供单个集成系统， 分离和检测。 最终，这将通过采样 毛细管到纳米管阵列，用于引入大量的 样品 该建议分为四个主要部分，包括： 优化分离DNA片段的双通道， 筛选缓冲液，2）开发改进的界面， 通过毛细管-通道界面的样品转移，3） Ru（bpy）32+作为电泳团的电化学阵列检测 dsDNA和ssDNA，以及4）毛细管转移方法的应用， 电化学阵列检测到芯片上的DNA分离。 这将 提供快速放置大量多个样品的方法 通道（1000个并行通道是可行的）在一个单一的芯片。 的 提出的技术有望将样品通量提高10至50倍 并将样本量减少2至3个数量级， 可用的系统。