Read e-book online DNA Viruses: Methods and Protocols PDF

By Feng Wang-Johanning, Gary L. Johanning (auth.), Paul M. Lieberman (eds.)

A compendium of conveniently reproducible and novel the way to manage DNA viruses and symbolize their various organic houses. The authors emphasize recommendations for viral detection and genetics, but additionally comprise tools for constitution selection, gene expression, replication, pathogenesis, complicated mobile versions, recombinant genetics, and computational/systems ways. Wide-ranging and hugely useful, DNA Viruses: tools and Protocols will stimulate new instructions in virology learn with its novel options for engineering viral vectors in gene remedy, and its complex methods for detecting viruses in human ailment.

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1, 75–82. 24 Stevens, Verkuijlen, and Middeldorp 4. 4 Yao, Q. , Rickinson, A. , and Epstein, M. A. (1985) A re-examination of the Epstein-Barr virus carrier state in healthy seropositive individuals. Int. J. Cancer. 35, 35–42. 5. 5 Stevens, S. J. , Verschuuren, E. A. , Verkuijlen, S. A. W. , van den Brule, A. J. , Meijer, C. J. L. , and Middeldorp, J. M. (2002) Role of Epstein-Barr virus DNA load monitoring in prevention and early detection of post-transplant lymphoproliferative disease. Leuk. Lymphoma 43, 831–840.

ZEBRA (primers Z1 and Z2). i. The cellular housekeeping gene U1A snRNP (primers U1A1 and U1A2). 2 µL AmpliTaq DNA polymerase. Multiprimed EBV RT-PCR 33 4. Cycle the samples in a PCR device using the following PCR program: 4 min at 95°C; 40 cycles of 1 min at 95°C, 1 min at 55°C, and 1 min at 72°C; 7 min at 72°C, and finally a hold at 4°C (see Note 15). 4. 1. Electrophoretic Separation of PCR Products and Blotting to Nylon 1. 5% agarose gel electrophoresis in TBE buffer for 1–2 h at approx 100 mA.

Equilibrate the column with 1 mL of TE before adding the end-labeling reaction mixture. Do not leave the column standing dry at any moment but continually keep adding 200-µL TE fractions. Collect fractions of 200 µL in different tubes for radioactivity measurements. 19. Hybridize and wash at exactly 55°C. Higher temperatures may lead to negative results, as the probe may be unable to bind and lower temperatures may increase nonspecific background hybridization. 20. Radiolabeled probes in hybridization buffer can be stored at –20°C for reuse.

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