Lipoproteins and Cardiovascular Disease, Softcover reprint of the original 1st ed. 2013 Methods and Protocols Methods in Molecular Biology Series, Vol. 1027

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.

Part I: RNA and Gene Expression

1. Cloning Full-Length Transcripts and Transcript Variants Using 5’ and 3’ RACE

            Lita A. Freeman

2. Monitoring Gene Expression: Quantitative Real-Time RT-PCR

            Elke M. Wagner

3. Microarray Technology: Basic Methodology and Application in Clinical Research for Biomarker Discovery in Vascular Diseases

            Nalini Raghavachari

4. Northern Analysis of Gene Expression

            Lita A. Freeman

5. Laser Capture Microdissection for Analysis of Macrophage Gene Expression from Atherosclerotic Lesions

            Jonathan E. Feig and Edward A. Fisher

Part II: Sequencing

6. Sequencing PCR-Amplified DNA in Lipoprotein and Cardiovascular Disease Research

            Victoria Youngblood and James G. Taylor VI

7. Introduction to Next-Generation Nucleic Acid Sequencing in Cardiovascular Disease Research

            Lena Diaw, Victoria Youngblood, and James G. Taylor VI

Part III: Transgenic, Knockout, and Knockdown Methodologies

8. Strategies for Designing Transgenic DNA Constructs

            Chengyu Liu

9. Purification of Plasmid and BAC Transgenic DNA Constructs

            Chengyu Liu, Yubin Du, Wen Xie, and Changyun Gui

10. Pronuclear Microinjection and Oviduct Transfer Procedures for Transgenic Mouse Production

            Chengyu Liu, Wen Xie, Changyun Gui, and Yubin Du

11. Genotyping of Transgenic Animals by Real-Time Quantitative PCR with TaqMan Probes

            Boris Vaisman

12. Generation of General and Tissue-Specific Gene Knockout Mouse Models

            Xian-Cheng Jiang

13. Adeno-Associated Viruses as Liver-Directed Gene Delivery Vehicles: Focus on Lipoprotein Metabolism

            William R. Lagor, Julie C. Johnston, Martin Lock, Luk H. Vandenberghe, and Daniel J. Rader

14. Modulation of Lipoprotein Metabolism by Antisense Technology: Preclinical Drug Discovery Methodology

            Rosanne M. Crooke and Mark J. Graham

Part IV: Special Topics

15. Chromatin Immunoprecipitation

            Grant D. Barish and Rajenda K. Tangirala

16. Measurement of Lecithin:Cholesterol Acyltransferase Activity with the Use of a Peptide-Proteoliposome Substrate

            Boris Vaisman and Alan T. Remaley

17. Native-Native 2D Gel Electrophoresis for HDL Subpopulation Analysis

            Lita A. Freeman

18. Western Blots

           Lita A. Freeman

Encompasses lipoprotein-centered methodologies designed for all experience levels

Features detailed protocols that are lab-ready

Includes expert tips and key implementation advice to ensure successful results