Adenovirus Methods and Protocols: Volume 1: Adenoviruses, Ad by William S. M. Wold, Ann E. Tollefson

By William S. M. Wold, Ann E. Tollefson

Adenovirus tools and Protocols, moment version, now in volumes, is an important source for adenovirus (Ad) researchers starting within the box, and an inspirational place to begin for researchers trying to department into new components of advert learn. as well as updating and increasing the 1st version, the authors have additional new chapters that handle leading edge parts of emphasis in advert study, together with advert vector development and use, real-time PCR, use of recent animal types, and strategies for quantification of advert virus or virus expression/interactions. all the protocols awarded in those volumes is written through trendsetting researchers.

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Extra resources for Adenovirus Methods and Protocols: Volume 1: Adenoviruses, Ad Vectors, Quantitation, and Animal Models (Methods in Molecular Medicine)

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0. Store at room temperature. 18. EcoRI (20 U/μL). 19. 1 M MgCl2, 10 mM dithiothreitol (DTT). Add DTT before using. Store at room temperature. 20. 100 mM DTT. Store at –20°C. 21. Klenow polymerase (5 U/μL). Store at –20°C. 22. 5. Store at –20°C. 23. 1 μg/μL Sheared salmon sperm DNA (29). Store at –20°C. 24. Mouse 3T6 cells (American Type Culture Collection). 25. 100X Glutamine: 29 g L-glutamine in 1 L of water. Filter-sterilize and aliquot. Store at –20°C long term or at 4°C while in use. 26. 100X Pen/strep: 10,000 U (Gibco 15140-122).

Dedieu, J. , and Perricaudet, M. (1996) Efficient dual transcomplementation of adenovirus E1 and E4 regions from a 293-derived cell line expressing a minimal E4 functional unit. J. Virol. 70, 559–565. 45. Brough, D. , Kulesa, V. , and Kovesdi, I. (1996) A gene transfer vector-cell line system for complete functional complementation of adenovirus early regions E1 and E4. J. Virol. 70, 6497–6501. 46. , Jia, X. , and Finer, M. H. (1995) A packaging cell line for propagation of recombinant adenovirus vectors containing two lethal gene-region deletions.

150-mm Polystyrene tissue culture plates (Costar 0877224). 71. Small glass test tubes with caps. 72. 6% Agarose: weigh agarose (low EEO). Add water and autoclave. Store at room temperature. 73. 8 g NaHCO3. 2 with HCl. Bring volume to 1700 mL. 2-μm filter to 425-mL aliquots. Incubate one bottle at 37°C overnight to check for contamination. Per 425 mL of 2X DMEM, add 10 mL 100X glutamine, 10 mL 100X pen/strep, 10 mL 100X nonessential amino acids, 25 mL 1 M MgCl2, and 20 mL HICS. When these components are added, the solution is 2X plaque assay medium.

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