I. Preparation of Virus Particles and Their Analysis 1. Methods for Viral RNA Isolation and PCR Amplification for Sequencing of Near Full-Length HIV-1 Genomes Kimdar Sherefa Kemal, Milan Reinis, Barbara Weiser, Harold Burger 2. Purification of HIV-1 Virions by Subtilisin Digestion or CD45 Immunoaffinity Depletion for Biochemical Studies David E. Ott 3. Calculating HIV-1 Infectious Titer Using a Virtual TCID50 Method Yong Gao, Immaculate Nankya, Awet Abraha, Ryan M. Troyer, Kenneth N. Nelson, Andrea Rubio, and Eric J. Arts II. Methods to Study HIV-1 Replication Section 1. Early Events 4. Cell-free Assays for HIV-1 Uncoating Christopher Aiken 5. Real-Time PCR Analysis of HIV-1 Replication Post Entry Events Jean L. Mbisa, Krista A. Delviks-Frankenberry, James A. Thomas, Robert J. Gorelick and Vinay K. Pathak 6. Analysis of 2-LTR Circle Junctions of Viral DNA in Infected Cells Dibyakanti Mandal and Vinayaka R. Prasad 7. HIV-1 Recombination: An Experimental Assay and a Phylogenetic Approach Michael D. Moore, Mario P.S. Chin, and Wei-Shau Hu 8. Methods of Preparation and Analysis of Intracellular Reverse Transcription Complexes Ariberto Fassati 9. Analysis of Viral and Cellular Proteins in HIV-1 Reverse Transcription Complexes by Co-immunoprecipitation Sergey N. Iordanskiy and Michael I. Bukrinsky 10. Isolation and Analysis of HIV-1 Preintegration Complexes Alan Engelman 11. Bisarsenical Labeling of HIV-1 for Real-Time Fluorescence Microscopy Nathalie J. Arhel and Pierre Charneau Section 2. Late Events 12. Methods for the Study of HIV-1 Assembly Abdul A. Waheed, Akira Ono and Eric O. Freed 13. Assembly of Immature HIV-1 Capsids using a Cell-Free System Jaisri R. Lingappa and Beth K. Thielen 14. Preparation of Recombinant HIV-1 Gag Protein and Assembly of Virus-like Particles in vitro Siddhartha A. K. Datta and Alan Rein 15. Methods for the Analysis of HIV-1 Nucleocapsid Protein Interactions with Oligonucleotides Andrew G. Stephen and Robert J. Fisher 16. Methods for Analysis of Incorporation and Annealing of tRNALys in HIV-1 Shan Cen, Fei Guo, and Lawrence Kleiman III. Specialized Approaches to Study HIV-1 Biology and Pathogenesis 17. Somatic Cell Genetic Analysis to Identify HIV-1 Host Restriction Factors Susana T. Valente and Stephen P. Goff 18. Rapid, Controlled and Intensive Lentiviral Vector-based RNAi Manuel Llano, Natassia Gaznick and Eric M. Poeschla 19. Reverse Two Hybrid Screening to Analyze Protein-protein Interaction of HIV-1 and Cellular Proteins Supratik Das and Ganjam V. Kalpana 20. Methods to Study Monocyte migration induced by HIV-infected cells Vasudev R. Rao, Eliseo Eugenin, Joan Berman and Vinayaka Prasad 21. Novel Mouse Models for Understanding HIV-1 Pathogenesis Aviva Joseph, Kaori Sango, Harris Goldstein IV. Immunological Studies of HIV Section 1. Mucosal Immunology 22. Mucosal Antibody Responses to HIV Zina Moldoveanu and Jiri Mestecky 23. Isolating Mucosal Lymphocytes from Biopsy Tissue for Cellular Immunology Assays Barbara L. Shacklett, J. William Critchfield and Donna Lemongello Section 2. Measuring T cell Responses via Flow Cytometry 24. Quantifying HIV-1-spec
Despite major advances in HIV treatment, many areas require more study, in order to create efficacious, potent antiretrovirals that can suppress viral load completely and durably without toxic side effects, to define unknown drug targets and fine-tune known targets, and to better understand the interplay between viral and host factors. In "HIV Protocols, Second Edition", expert researchers provide clear, state-of-the-art methods for the study of HIV. Directed toward three specific goals, this text aims to document up-to-date protocols for select aspects of HIV biology, to bring together both virological and immunological approaches in a single, convenient volume, and to present a comprehensive account of a range of techniques not available in any existing HIV protocol book. As a volume in the highly successful Methods in Molecular Biology(TM) series, the chapters include brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections containing priceless tips on troubleshooting and avoiding known pitfalls.
Comprehensive and cutting-edge, "HIV Protocols, Second Edition" is an ideal guide to the wide array of techniques used in fundamental or applied research into the biology and pathogenesis of HIV-1.
Comprehensive account of cutting-edge, virological techniques with a wide range of protocols
Written by well-known HIV researchers selected for their expertise in the respective methodologies
Covers both virological and immunological techniques related to HIV in a single volume
Conveniently arranged protocols studying different aspects of viral replication reflect the order in which the events take place in the viral life cycle
Provides a breadth of coverage of many critical steps in HIV replication that could serve as targets for drug therapy