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控仪器适应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/取证应用的仪器的稍作修改的版本开始。我们希望利用现有的并行微流控和中观格式扫描共聚焦检测的新方面，在单人生物实验室的环境中为常规细胞分析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方面，该平台的表现优于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，该系统的吞吐量足以支持一个主要的克隆屏幕。这项开发将与怀特黑德研究所(Loish和Weinberg)的邻近实验室以及Wittrup的麻省理工学院实验室和布林赫斯特的MGH实验室合作进行。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。作为中等风险的延伸，我们将通过开发一种方法来演示深度表观遗传学分析，该方法通过对图像选择的细胞片段进行逆转录-聚合酶链式反应(RT-PCR)读出高度多元化(50-100个基因)的表达水平。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个通道的并行中完成。其目标是为生物学提供一个强大而多功能的工具。

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.