Download Hepatitis B and D Protocols: Volume 1: Detection, Genotypes, by Robert K. Hamatake, Johnson Y. N. Lau PDF

By Robert K. Hamatake, Johnson Y. N. Lau

A broad-ranging choice of center ideas for the learn of HBV and HDV infections and for the advance of remedies to regard them. during this first of 2 volumes Detection, Genotypes, and Characterization, the authors specialise in comfortably reproducible molecular tools for the identity and quantification of viral markers, the detection and impression of viral versions, and the research of the viral existence cycle. every one totally demonstrated protocol is defined in step by step element by means of a longtime professional within the box and features a history advent outlining the main underlying the method, gear and reagent lists, and pointers on troubleshooting and fending off recognized pitfalls. An accompanying moment quantity, Immunology, version structures, and scientific stories, includes elementary protocols for the research of host immune responses to an infection, in vitro and in vivo types of an infection, and the improvement of antivirals. For either volumes: A broad-ranging choice of center recommendations for the research of HBV and HDV infections and for the improvement of remedies to regard them. the 1st quantity Detection, Genotypes, and Characterization, the authors specialize in comfortably reproducible molecular tools for the id and quantification of viral markers, the detection and impression of viral editions, and the examine of the viral existence cycle. the second one quantity, Immunology, version platforms, and scientific stories, comprises uncomplicated protocols for the examine of host immune responses to an infection, in vitro and in vivo versions of an infection, and the advance of antivirals. each one absolutely proven protocol is defined in step by step element by means of a longtime specialist within the box and features a history advent outlining the main underlying the process, gear and reagent lists, and tips about troubleshooting and fending off recognized pitfalls. The set deals either new and skilled investigators an encyclopedic selection of strong instruments for learning HBV and HDV infections, a necessary source for locating new remedies to regard chronically contaminated sufferers.

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Additional info for Hepatitis B and D Protocols: Volume 1: Detection, Genotypes, and Characterization

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Serum or plasma samples containing HBsAg, 10 ␮L per run. 6. For dilutions: negative human or fetal calf serum or plasma. Do not mix serum with plasma. 2. Equipment and Disposables 1. Glass slides 2 × 3 in. 5 cm) with smooth edges to avoid cuts. Glass slides should be fat free. 1% hot agarose solution in hot water. 60 Gerlich, Wend, and Glebe Fig. 1. The electrophoresis chamber and the gel punch built at the workshop of the Institute of Medical Virology. 2. , Schleicher and Schuell cat. no. 2668/8).

5 ␮L (100 ng/␮L) each of the primers 1445+ and 1574− is added, with a final volume of 50 ␮L. 3. Incubate tubes in a thermocycler at 50°C for 20 min (reverse transcription). 4. Subsequent steps in the cycler correspond to the PCR protocol as described above. 5. PCR and RT–PCR can also be carried out using the primer pair 1434+ and 1668−, resulting in the amplification of a larger segment shared by both f and tr transcripts. Here, the annealing temperature is 56°C. 3. f RNA Assay RT–PCR involving anchored oligo (dT) primers recognizing full-length transcripts terminating at the polyadenylation signal downstream of the X reading frame is carried out in two rounds.

69, 3350–3357. 3. , and Mason, W. S. (1982) Replication of the genome of a hepatitis B-like virus by reverse transcription of an RNA intermediate. Cell 29, 403–415. 4. Tuttleman, J. , and Summers, J. (1986) Formation of the pool of covalently closed circular viral DNA in hepadnavirus-infected cells. Cell 47, 451–460. 5. Jilbert, A. , Wu, T. , England, J. , et al. (1992) Rapid resolution of duck hepatitis B virus infections occurs after massive hepatocellular involvement. J. Virol. 66, 1377–1388.

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