Fluidigm BioMark High-Throughput Genetic Analysis System

描述（由申请人提供）：基因表达分析彻底改变了我们对生物学基本问题和临床疾病研究的理解。研究基因表达的两种传统方法是在有限条件下进行高通量研究的微阵列分析和对少数基因进行精确和敏感定量的实时聚合酶链反应（PCR）。微阵列分析相对缺乏灵敏度和高昂的费用是限制发现生物过程中基因表达变化的主要瓶颈，也阻碍了系统生物学方法对基因调控的研究。与传统实时PCR分析相关的高试剂成本和费力的样品处理通常妨碍了使用这种强大的方法进行高通量分析。精确控制纳升液体体积的微流控技术的发展极大地提高了实时PCR研究的潜在通量。最先进的BioMark高通量遗传分析系统使用集成流体电路（IFCs），以极低的成本将实时PCR的精度和灵敏度与极高的通量结合起来。此外，该系统能够以适合大规模群体遗传学研究的通量进行单核苷酸多态性（SNP）基因分型，高通量拷贝数变异分析和数字PCR应用，提供单细胞基因表达的高定量分析。由13名美国国立卫生研究院资助的宾夕法尼亚大学医学院、艺术与科学学院和天普大学的研究人员组成的联盟，他们的研究得到了68项美国国立卫生研究院的资助，他们将成为所要求的生物标记系统的主要用户，该系统将被安置在宾夕法尼亚大学微阵列核心设施中。该设备应用程序支持的项目将利用生物标记系统的功能来研究宾夕法尼亚大学目前可用的微阵列和低通量实时PCR系统无法解决的关键行为和生物医学问题。这些研究有望彻底改变我们对昼夜节律、记忆储存、睡眠、精神疾病、代谢调节和心血管功能背后的生物过程的理解，为未来开发针对各种疾病的新治疗方法奠定基础。

DESCRIPTION (provided by applicant): The analysis of gene expression has revolutionized our understanding of fundamental questions in biology and the study of clinical disorders. The two traditional approaches to study gene expression are microarray analysis for high-throughput studies under a limited set of conditions and real-time polymerase chain reaction (PCR) for precise and sensitive quantification of a few genes. The relative lack of sensitivity and high expense in microarray analysis are major bottlenecks limiting the discovery of gene expression changes during biological processes and hindering systems-biological approaches to gene regulation. The high reagent costs and laborious sample handing associated with traditional real-time PCR analysis have generally precluded high-throughput analysis using this otherwise powerful approach. The development of microfluidic technologies to precisely manipulate nanoliter liquid volumes has dramatically increased the potential throughput of real-time PCR studies. The state-of-the-art BioMark High-Throughput Genetic Analysis System uses integrated fluidics circuits (IFCs) to combine the precision and sensitivity of real-time PCR with extremely high-throughput at a fraction of the costs. Further, this system is capable of single nucleotide polymorphism (SNP) genotyping at a throughput amenable to large-scale population genetics studies, high-throughput copy number variation analysis, and digital PCR applications that provide highly quantitative analysis of gene expression from single cells. A consortium of 13 NIH-funded researchers at the University of Pennsylvania School of Medicine, School of Arts and Sciences and Temple University, whose research is supported by 68 NIH grants, will be major users of the requested BioMark system, which will be housed in the University of Pennsylvania Microarray Core Facility. The projects supported by this equipment application will take advantage of the capabilities of the BioMark System to study critical behavioral and biomedical questions that cannot be addressed with the currently available microarray and low throughput real- time PCR systems at the University of Pennsylvania. These studies promise to revolutionize our understanding of the biological processes underlying circadian rhythms, memory storage, sleep, psychiatric disease, metabolic regulation and cardiovascular function, laying the groundwork for the future development of novel therapeutic approaches to a variety of disorders.