Analytical Core

分析中心将为计划项目的研究人员提供各种专业的分析服务，以协助进行分子、细胞和体内研究。分析核心将是在加州大学圣地亚哥分校的约瑟夫·维茨图姆博士和索尔克研究所的罗纳德·埃文斯博士指导下的一个单位之间的合作努力。分析核心的总体目标是利用专业资源和专门知识，为研究人员提供选定的核心服务，以协助每个项目的研究使命。该核心的目标是： 具体目标1：提供基因特征和途径的高通量纳米级定量PCR分析（Nano-QPCR）分析。在这里，我们描述了高通量纳米级定量，实时，逆转录PCR（Nano-QPCR）的设备和操作程序，用于调查SNP基因分型和分析PPG研究人员提供的样品中特定转录本的表达。 具体目标2：使用Luminex Bio-PCR技术提供高通量、高灵敏度的细胞因子和趋化因子分析。核心的Bio-Biological悬浮阵列组件已经在Evans实验室运行了10多年，由Luminex Bio-Biological工作站组成，该工作站将促进细胞因子和趋化因子在足够小的样品体积中的蛋白质组学分析，该样品体积与血浆或培养液以高通量格式兼容。 具体目标3：为PPG研究者提供免疫学支持。免疫学核心将生成针对所需抗原的抗血清，并根据要求制备供其他项目使用的各种免疫学试剂，包括用于免疫测定或Western印迹或FACS分析的一抗或二抗。 具体目标4：提供脂质和脂蛋白水平的定量和定性分析，并提供脂蛋白组分。核心将提供总胆固醇和甘油三酯的测量，并将分析这些分析物的鼠血浆FPLC谱，并根据需要提供人LDL和/或其他脂蛋白

The Analytical Core will provide a variety of specialized analytical services for the investigators of the Program Project to assist in the conduct ofthe molecular, cellular and in vivo studies. The Analytical Core will be a collaborative effort between units at the University of California, San Diego under Dr. Joseph Witztum and one at the Salk Institute under the direction of Dr. Ronald Evans. The overall goal of the Analytical Core is to take advantage of specialized resources and expertise to provide investigators with selected core services that will assist in the research mission of each Project. The Aims of this Core are: Specific Aim 1: To provide high-throughput nano-scale quantitative PCR analysis (Nano-QPCR) profiling of gene signatures and pathways. Here we describe the equipment and operation procedures for high-throughput nano-scale quantitative, real-time, reverse-transcription PCR (Nano-QPCR) for surveying SNP genotyping and profiling the expression of particular transcripts in samples provided by PPG investigators. Specific Aim 2: To provide high throughput, high sensitivity analysis of cytokines and chemokines with the use of Luminex Bio-Plex technology. The Bio-Plex suspension array component ofthe core has been in operation in the Evans laboratory for over 10 years and consists of a Luminex Bio-Plex workstation that will facilitate proteomic profiling of cytokines and chemokines in sufficiently small sample volumes compatible with plasma or culture fluids in a high throughput format. Specific Aim 3: To provide immunological support for investigators of the PPG. The Immunology Core will generate antisera against desired antigens and prepare a wide variety of immunological reagents to be used by the other Projects as requested, including primary or secondary antibodies to be used in immunoassays or Western blots or FACS analysis. Specific Aim 4: To provide quantitative and qualitative analysis of lipid and lipoprotein levels and provision of lipoprotein fractions. The core will provide measures of total cholesterol and triglycerides and will analyze FPLC profiles of murine plasma for these analytes and provide human LDL and/or other lipoproteins as needed