X, 391 p. 50 illus., 23 illus. in color. online resource
SUMMARY
The lipid-rich and otherwise challenging nature of many key tissues complicates many aspects of current research, and applications of the unique nature of lipoproteins and their biological effects has engendered unique and vital methodologies. In Lipoproteins and Cardiovascular Disease: Methods and Protocols, experts in the field present a compendium of advanced and classical molecular biology methods targeted towards lipoprotein, atherosclerosis, and vascular biology research, bringing together in a single volume an updated set of protocols and strategies for methods now driving the most recent advances, along with classical methods that are still widely used. Among the many topics covered in this cutting-edge work, the book delves into crucial techniques such as quantitative real-time PCR, microarrays, RT-PCR laser capture microdissection, and tissue-specific gene overexpression, knockout, and knockdown methodologies, including AAV as a liver-directed gene delivery vehicle. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and valuable notes which highlight tips on troubleshooting and avoiding known pitfalls. Comprehensive and easy to use, Lipoproteins and Cardiovascular Disease: Methods and Protocols serves both novices and experts alike as a complete guide for any researcher with an interest in lipoproteins and their significant biological effects
CONTENT
Cloning Full-Length Transcripts and Transcript Variants Using 5’ and 3’ RACE -- Monitoring Gene Expression: Quantitative Real-Time RT-PCR -- Microarray Technology: Basic Methodology and Application in Clinical Research for Biomarker Discovery in Vascular Diseases -- Northern Analysis of Gene Expression -- Laser Capture Microdissection for Analysis of Macrophage Gene Expression from Atherosclerotic Lesions -- Sequencing PCR-Amplified DNA in Lipoprotein and Cardiovascular Disease Research -- Introduction to Next-Generation Nucleic Acid Sequencing in Cardiovascular Disease Research -- Strategies for Designing Transgenic DNA Constructs -- Purification of Plasmid and BAC Transgenic DNA Constructs -- Pronuclear Microinjection and Oviduct Transfer Procedures for Transgenic Mouse Production -- Genotyping of Transgenic Animals by Real-Time Quantitative PCR with TaqMan Probes -- Generation of General and Tissue-Specific Gene Knockout Mouse Models -- Adeno-Associated Viruses as Liver-Directed Gene Delivery Vehicles: Focus on Lipoprotein Metabolism -- Modulation of Lipoprotein Metabolism by Antisense Technology: Preclinical Drug Discovery Methodology -- Chromatin Immunoprecipitation -- Measurement of Lecithin:Cholesterol Acyltransferase Activity with the Use of a Peptide-Proteoliposome Substrate -- Native-Native 2D Gel Electrophoresis for HDL Subpopulation Analysis -- Western Blots
