By Feng Wang-Johanning, Gary L. Johanning (auth.), Paul M. Lieberman (eds.)
A compendium of comfortably reproducible and novel how you can manage DNA viruses and represent their diversified organic homes. The authors emphasize options for viral detection and genetics, but additionally comprise tools for constitution selection, gene expression, replication, pathogenesis, advanced mobile versions, recombinant genetics, and computational/systems ways. Wide-ranging and hugely sensible, DNA Viruses: tools and Protocols will stimulate new instructions in virology examine with its novel options for engineering viral vectors in gene remedy, and its complex ways for detecting viruses in human affliction.
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Extra info for DNA Viruses: Methods and Protocols
Sample text
The cycle at which the fluorescence exceeds the background is designated the threshold cycle. By comparing the threshold cycle of a clinical specimen with those of standard curve samples, the amount of EBV DNA in clinical samples can be determined. This real-time PCR approach is extremely rapid owing to efficient heat conduction by using glass capillaries, small reaction volumes, and air as heating medium. The “closed-tube” system eliminates the risk of PCR contamination by product carryover and also the need for post-PCR detection.
EBNA-2 (primers EBNA-2 Y2 and EBNA-2 H1). d. 2). e. LMP-2a (primers LMP-2a1 and LMP-2ab2). f. LMP-2b (primers LMP-2b1 and LMP-2ab2). g. BARTs (primers A3 and A4). h. 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).
M. , Totowa, NJ 15 16 Stevens, Verkuijlen, and Middeldorp ing number of malignant proliferative disorders of both lymphoid and epithelial origin, including Burkitt’s and Hodgkin’s lymphoma, B- and T-cell nonHodgkin’s lymphoma, and nasopharyngeal and gastric carcinoma (1,2). In immunocompromised individuals such as transplant recipients and AIDS patients, EBV is the driving force behind lymphoproliferative disorders, originating from infected B cells, which initially are benign and polyclonal but may progress to malignant lymphoma if left untreated (1).