Roche Real-Time Polymerase Chain Reaction Workflow System

描述(由申请人提供)：我们要求一个罗氏实时聚合酶链式反应(PCR)工作流程系统，具有组织制备(Magna Lyser)、核酸分离(Magna Pure Comp)和实时PCR(LightCycler 480)的自动化组件。LightCycler 480是一种检测和量化核酸序列的高通量实时聚合酶链式反应仪器，具有96孔板和384孔板的容量。这套聚合酶链式反应工作流程系统非常易于使用，并配有功能强大的软件，使研究人员能够在3-4小时内从组织到核酸定量。该仪器将被安置在博斯沃思健康科学研究大楼的大学微阵列核心研究设施中，并将由毒理学研究生院以及解剖学和神经生物学系的教职员工使用。参与这一申请的研究人员小组包括七名拥有NIH资助的资助项目的教师，他们是主要用户，以及另外三名次要用户的教师。研究项目多种多样，但主要围绕三个主题：1)氧化应激导致的组织损伤和癌症，2)DNA损伤、修复和突变，以及3)基因表达的激素调节。我们打算使用该仪器对生物样本中的RNA转录本和DNA靶标进行高通量实时定量分析，并检测在癌症中重要的基因的单核苷酸多态(SNP)。正如每个调查员的研究项目所表明的那样，拟议的工具的提供将大大改善他们研究的范围和及时性。最重要的是，它将允许科学界决定在其资助的研究项目中追求令人兴奋的发现，而不是现有仪器的可用性。这一共享的多学科研究机构的总体目标是利用这一先进技术资源开展具有当今生物医学研究特点的尖端研究，并培训我们的博士生和博士后使用这些现代方法，使他们能够发展成为成功的独立研究人员。我们的研究人员所解决的研究问题与与癌症有关的重大公共卫生问题高度相关，无论是在肿瘤形成的机制、癌症化疗药物(缺乏)有效性的原因，还是对女性健康至关重要的激素对基因的调节方面。

DESCRIPTION (provided by applicant): We are requesting a Roche real-time polymerase chain reaction (PCR) Workflow System with automated components for preparation of tissues (MagNA Lyser), isolation of nucleic acids (MagNA Pure Compact), and a real-time PCR (LightCycler 480). The LightCycler 480 is a high throughput real- time PCR instrument that detects and quantifies nucleic acid sequences, and has a 96- and 384-well plate capacity. This PCR Workflow System is very easy to use, is accompanied by powerful software, and enables an investigator to go from tissue to nucleic acid quantitation in 3-4 hours. This instrument will be housed in the MicroArray Core Research Facility of the University in the Bosomworth Health Sciences Research Building, and will be used by faculty in the Graduate Center for Toxicology, and in the Department of Anatomy and Neurobiology. The group of investigators participating in this application includes seven faculty with NIH-supported grant projects who are major users, and three additional faculty who are minor users. The research projects are diverse, but revolve around three major themes: 1) oxidant-stress induced tissue injury and cancer, 2) DNA damage, repair and mutagenesis and 3) hormonal regulation of gene expression. We intend to use this instrument for the high-throughput real-time quantitative analysis of RNA transcripts and DNA targets in biological samples and in detection of single nucleotide polymorphisms (SNP) in genes important in cancer. As indicated in the research projects from each investigator, the availability of the proposed instrument will significantly improve the scope and timeliness of their studies. Most importantly, it will allow the science to determine the pursuit of exciting discoveries in their funded research projects, rather than availability of current instruments. The overall goals of this shared multidisciplinary research facility are to use this advanced technology resource to carry out cutting-edge research that characterizes today's biomedical research and to train our doctoral students and postdoctoral fellows in the use of these modern methods that will enable them to develop into successful independent investigators. The research problems addressed by our investigators are highly relevant to major public health problems related to cancer, both in the mechanisms of tumor formation, the reasons for (lack of) effectiveness of cancer chemotherapeutic drugs, and the regulation of genes by hormones that are important in women's health.