Description
Lipoproteins and Cardiovascular Disease, Softcover reprint of the original 1st ed. 2013
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1027
Coordinator: Freeman Lita A.
Language: EnglishSubjects for Lipoproteins and Cardiovascular Disease:
Support: Print on demand
391 p. · 17.8x25.4 cm · Hardback
Description
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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.
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
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