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
The purpose of this core is to provide analytical expertise to the PPG investigators by making available specialized resources that will greatly facilitate the conduct of research. As specific questions are developed, targeted use of techniques available in this core can be developed.
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