High Throughput BioMEMS DNA Sequencing

描述（由申请人提供）：我们将开发桥接技术，使我们的手动和自动微流体仪器适应于基于细胞的分析应用。我们的目标是从我们已经商业化的用于DNA测序/法医应用的仪器的稍微修改版本开始。我们希望利用现有的平行微流体和中观格式扫描共聚焦检测的新方面，在单个研究人员的生物实验室环境中为常规细胞检测创造新的低成本仪器。大框架BioMEMS-768 DNA测序仪成为自动化平台的起点，我们将证明，在一类稀有细胞，原代细胞和定量标记应用中优于FACS。桌面大小的GeneTrack系统将成为廉价的半手动系统的基础，尽管如此，它有足够的吞吐量来支持一个主要的克隆屏幕。该开发将与Whitehead研究所（Lodish和Weinberg）的邻近实验室、Wittrup的麻省理工学院实验室和Bringhurst的MGH实验室合作。这些合作者将通过我们的仪器进行大规模的克隆筛选和稀有细胞分析。作为中等风险的扩展，我们将通过开发一种在图像选择的细胞片段上通过反转录PCR读取高度多路（50-100个基因）表达水平的方法来展示深度表观遗传学分析。这一切都将通过一种新的实时RT-PCR电泳多路复用方法在384车道并行中完成。目标是为生物学提供一个强大的多功能工具。

DESCRIPTION (provided by applicant): We will develop the bridging technology to adapt our manual and automated microfluidic instruments to applications in cell-based assays. Our goal is to start with only slightly modified versions of instruments that we have already commercialized for DNA sequencing/forensics applications. We hope to exploit novel aspects of the existing parallel microfluidics and meso-format scanning confocal detection, to create new low-cost instruments for routine cell-based assays in the environment of the single-investigator biology lab. The large-frame BioMEMS-768 DNA sequencer becomes the starting point for an automated platform that, we will prove, outperforms FACS for a class of rare-cell, primary-cell and quantitative-marker applications. The desk-top-sized GeneTrack system will be the basis for an inexpensive semi-manual system, which, nonetheless, has sufficient throughput to support a major cloning screen. The development will be collaborative with neighboring laboratories at the Whitehead Institute (Lodish and Weinberg), as well as an MIT Laboratory of Wittrup, and the MGH laboratory of Bringhurst. These collaborators will run large-scale cloning screens and rare-cell assays though our instruments. As a moderate-risk extension, we will demonstrate deep epigenetic analysis by developing a means to read out highly multiplexed (50-100 gene) expression levels by reverse-transcription PCR on image-selected cell fractions. This will all be done in 384-lane parallelism by a novel real-time RT-PCR electrophoretic multiplexing method. The goal is to provide a powerful and multifunctional tool for biology.