AUTOMATED HIGH SPEED DNA SEPARATIONS

描述：开发高通量DNA采样、分离和 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、小容量DNA的新技术 分居。这将通过结合毛细管样品来完成 在微细加工芯片中引入并行分离，以及 电化学阵列检测，以提供单个集成系统 分离和检测。最终，这将通过抽样将其联系起来 毛细管到纳米小瓶阵列，用于引入大量的 样本。该提案分为四大部分，包括1) 超薄通道中DNA片段分离的优化 筛选缓冲器，2)开发改进的接口以允许高效 通过毛细管-通道界面的样品转移，3)发展 以Ru(Bpy)32为电泳液的电化学阵列检测 DsDNA和ssDNA，以及4)毛细管转移法和 芯片上DNA分离的电化学阵列检测。这将是 提供快速大量放置多个样品的方法 在单个芯片上实现多个通道(1000个并行通道是可行的)。这个 拟议的技术有望将样品吞吐量提高10到50倍 与目前相比，样本量减少了2到3个数量级 可用的系统。

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.