Download HIV Protocols by Vinayaka R. Prasad, Ganjam V. Kalpana (eds.) PDF

By Vinayaka R. Prasad, Ganjam V. Kalpana (eds.)

This 3rd variation quantity expands upon the former variations with new learn issues and diverse modern HIV protocols. This booklet comprises chapters that conceal a variety of features of HIV-1 replication and are beneficial for investigating the mechanisms of HIV-1 replication or pathogenesis. The chapters are grouped into six sections: HIV early occasions; HIV-1 RNA constitution and RNA-protein interactions; HIV-1 post-integration occasions; HIV-1 pathogenesis in animal versions; instruments to review HIV-1 latency; and Neuro AIDS. Chapters are written by means of study specialists who've diligently built recommendations, superior them, simplified them, or utilized them to their very own HIV study. Written within the hugely profitable Methods in Molecular Biology series layout, the chapters comprise introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, effectively reproducible laboratory protocols and tips about troubleshooting and keeping off identified pitfalls.

Authoritative and state of the art, HIV Protocols, 3rd Edition is a brilliant source for HIV learn, and includes many major tips and methods not often pointed out in normal protocols.

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Chikere K, Webb NE, Chou T, Borm K, Sterjovski J, Gorry PR, Lee B (2014) Distinct HIV-1 entry phenotypes are associated with transmission, subtype specificity, and resistance to broadly neutralizing antibodies. Retrovirology 11:48. 1186/1742-4690-11-48 Chapter 2 Measuring T Cell-to-T Cell HIV-1 Transfer, Viral Fusion, and Infection Using Flow Cytometry Natasha D. Durham and Benjamin K. Chen Abstract Direct T cell-to-T cell HIV-1 infection is a distinct mode of HIV-1 infection that requires physical contact between an HIV-1-infected “donor” cell and an uninfected, CD4-expressing “target” cell.

The cell seed provided is optimized for luciferase reporter pseudotype virus requiring a 48 h postinfection culture period. Longer culture periods will require lower cell seeds to prevent overgrowth. The ultimate goal is to achieve a healthy and adherent cell density on the day of infection that is low enough to prevent overgrowth until infection is measured. Seeding Affinofile cells onto polylysine-coated plates can result in less well-to-well variability, especially when highly passaged Affinofile cells begin to lose their already weak baseline adherence.

5. Resuspend target cells in the appropriate volume of 6 µM Cell Proliferation dye eFluor 450. Resuspend donor cells in the appropriate volume of 4 µM Cell Proliferation dye eFluor 670. 6. Incubate cells at 37 °C for 10 min in the dark. 7. Add 4–5 volumes of RPMI complete to stop the labeling reaction. Spin at 400 × g for 5 min. 8. Remove supernatant and resuspend cells in 4–5 volumes of RPMI complete. Spin at 400 × g for 5 min to wash cells (wash #1). 9. Remove supernatant and resuspend cell pellet in 1 or 2 mL of RPMI complete.

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