Molecular Biology Core Laboratory

分子生物学核心实验室

基本信息

批准号：
8106525
负责人：
DAVID W. RUSSELL
金额：
$ 73.51万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8106525
关键词：
Acrylamides Actins Address Agar Gel Electrophoresis Age Aliquot Amino Acid Sequence Archives Arterial Fatty Streak Arteries Atherosclerosis Autoradiography Biological Biotin Blood Blood Vessels Blood specimen Brain Budgets Buffers Cataloging Catalogs Caveolae Cell Proliferation Cell surface Cells Cellular biology Charge Chromatography Chromosome Mapping Collection Communities Complementary DNA Complementary RNA Computer software Contracts Core Facility Custom Cyclophilins DNA DNA Binding DNA Sequence DNA Sequencing Facility Data Databases Deposition Development Digestion Electronic Mail Electrophoresis Endocytosis Endopeptidase K Funding Future Gel Gene Expression Gene Family Gene Proteins Gene Structure Genes Genetic Genetic Engineering Genomic Library Genomics Genotype Glyceraldehyde 3-Phosphate Glycerol Goals Hour Human Human Resources Immunoblotting Immunoprecipitation Individual Institution Knock-out Knockout Mice Label Laboratories Length Libraries Los Angeles Low Density Lipoprotein Receptor Mails Maintenance Measures Mediation Membrane Messenger RNA Methods Modeling Molecular Biology Molecular Genetics Mus Oligonucleotide Microarrays Oligonucleotide Primers Oligonucleotides Oxidoreductase Pathology Pattern Peptide Sequence Determination Pharmacologic Substance Plasmids Platelet-Derived Growth Factor Receptor Polyacrylamide Gel Electrophoresis Polymerase Chain Reaction Population Power Sources Preparation Price Primer Extension Procedures Process Program Research Project Grants Property Proteins Protocols documentation RNA RNA Probes Reaction Receptor Gene Receptor Protein-Tyrosine Kinases Recombinant Proteins Recombinants Regulation Research Research Personnel Research Project Grants Reverse Transcription Ribosomal Proteins Robot Robotics Roentgen Rays Role Running Sampling Scanning Sequence Analysis Services Signal Pathway Signal Transduction Site Smooth Muscle Myocytes Sodium Dodecyl Sulfate-PAGE Specimen Synapses System Tail Techniques Technology Time Tissue Extracts Tissue-Specific Gene Expression Tissues Transgenes Transgenic Mice Transgenic Organisms Variant Work arm atherogenesis base cDNA Library cost design experience gel electrophoresis genetic analysis genetic pedigree insight laboratory facility lipid metabolism meetings mutant neurotransmission novel operation polyacrylamide gels programs protein purification research study restriction enzyme satisfaction sex tissue/cell culture

项目摘要

The Molecular Biology Core Laboratory is directed by Dr. Russell. He will be assisted in day-to-day operations by Dr. Jonathan Cohen, who is an expert in high-throughput DMA sequencing. This Core laboratory provides support for acrylamide gel electrophoresis of proteins, DMA sequencing, analysis of gene expression by oligonucleotide microarray hybridization and real-time polymerase chain reactions (PCRs), oligonucleotide procurement, genomic DMA and RNA isolation, and the maintenance and storage of bacterial strains, plasmids, and purified proteins used within this Program Project. Four experienced technicians, Kevin Anderson (100% time), Emily Brown (100%), Jeffrey Cormier (100%), and Scott Clark (25%) will perform the duties associated with this Core. The laboratory facility is located within the Department of Molecular Genetics. For the analysis of proteins, 30 vertical electrophoresis units and 12 multi-outlet power supplies are available. SDS polyacrylamide gels are either prepared (25% of gels) or purchased pre-poured (75% of gels) and run by a single technician (Emily Brown). Following electrophoresis, individual Investigators working on the different Research Projects process the gels for autoradiography, immunoblotting, or protein sequencing. A darkroom that contains a Konica automatic X-ray developer is used to process all autoradiograms and chemilumigrams. The services of the facility are used extensively to assess protein purification, immunoprecipitation from cells, expression of recombinant proteins, and immunoblotting from cultured cells, tissues, and recombinant hosts. These techniques and methods are crucial to our studies on the expression and purification of essentially all genes and proteins with which we work. In addition, the determination of tissue-specific expression patterns of genes under study via immunoblotting in transgenic and knockout mice is heavily dependent on this aspect of the Core.

分子生物学核心实验室由Russell 博士领导。高通量 DMA 测序专家 Jonathan Cohen 博士将协助他进行日常操作。该核心实验室为蛋白质丙烯酰胺凝胶电泳、DMA 测序、通过寡核苷酸微阵列杂交和实时聚合酶链式反应 (PCR) 进行基因表达分析、寡核苷酸采购、基因组 DMA 和 RNA 分离以及细菌的维护和储存提供支持。本计划项目中使用的菌株、质粒和纯化蛋白质。四名经验丰富的技术人员 Kevin Anderson（100% 时间）、Emily Brown（100%）、Jeffrey Cormier（100%）和 Scott Clark（25%）将履行与该核心相关的职责。该实验室设施位于分子遗传学系内。对于蛋白质分析，可使用 30 个垂直电泳装置和 12 个多插座电源。 SDS 聚丙烯酰胺凝胶要么是制备的（25% 的凝胶），要么是购买的预浇注的（75% 的凝胶），并由一名技术人员 (Emily Brown) 运行。电泳后，从事不同研究项目的各个研究人员处理凝胶以进行放射自显影、免疫印迹或蛋白质测序。配备柯尼卡自动 X 射线显影机的暗室用于处理所有放射自显影图和化学发光图。该设施的服务广泛用于评估蛋白质纯化、细胞免疫沉淀、重组蛋白表达以及培养细胞、组织和重组宿主的免疫印迹。这些技术和方法对于我们研究几乎所有基因和蛋白质的表达和纯化至关重要。此外，确定通过转基因和基因敲除小鼠中的免疫印迹研究的基因的组织特异性表达模式在很大程度上取决于核心的这一方面。