Titlebook: DNA Viruses; Methods and Protocol Paul M. Lieberman Book 2005 Humana Press 2005 DNA.PCR.Termination.cell lines.electron microscopy.gene exp

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Viral Detectionain reaction (RT-PCR), PCR, real-time RT-PCR, and real-time PCR methods. Amplification of the E6 and E7 oncoproteins of HPV16 is described in detail, and primers and probes that can be used to amplify these oncogenes are described. Techniques to quantify these oncogenes in infected human tissue spec

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Quantitative Detection of Epstein-Barr Virus DNA in Clinical Specimens by Rapid Real-Time PCR Target, or plasma. This assay is based on amplification of a highly conserved 213-bp region of the . gene, a single-copy gene of EBV required for maintenance of the EBV genome within the infected host cell. For real-time detection of amplicons, two internal hybridization probes are added, labeled with the

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Profiling of Epstein-Barr Virus Latent RNA Expression in Clinical Specimens by Gene-Specific Multiprrus. As a control for RNA integrity, the low-copy-number transcript derived from U1 A snRNP, a cellular housekeeping gene, is coamplified. Copy DNA (cDNA) for these nine targets is simultaneously synthesized in a gene-specific, multiprimed cDNA reaction, which strongly reduces the amount of required

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Quantitative Detection of Viral Gene Expression in Populations of Epstein-Barr Virus-Infected Cells ected cells within a given population are expressing the viral genes . Q-K, ., ., ., ., and the .. Because this technique involves limiting dilution analysis, it is possible to define which viral transcription programs are being used at the single-cell level. This assay takes 3-4 d to complete and i

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Detection and Quantification of the Rare Latently Infected Cell Undergoing Herpes Simplex Virus Tranm latency in a number of nonhuman species, including mice. This provides a unique opportunity to study the complex lytic-latent cycle of a human neurotropic virus in a mouse model. This chapter details basic methods for inducing and quantifying reactivation, with emphasis on the first strategy for d

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Reporter Cell Lines for the Detection of Herpes Simplex Virusesechniques. However, virus culture is generally a slower process, as it inevitably takes the period of a full replication cycle of a given virus. A genetically modified cell culture with a virus-inducible marker is described here, using a frequently isolated DNA virus (herpes simplex virus) as a mode

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Studying the Structure of Large Viruses With Multiresolution Imagingirements of protein crystallography. The methodology consists in fitting the atomic structures of capsid components, independently solved, to medium-resolution, 3D maps of the complete virion, obtained by cryoelectron microscopy and image processing. On combining the two kinds of imaging data, one m

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Herpes Simplex Virus-Cell Interactions Studied by Low-Fading Contrasted Immunofluorescence other cell biology studies because: (.) it is a simple, rapid, sensitive, and reproducible technique; (.) phase-contrast microscopy is unnecessary; (.) contrast is optimal without blurring the fluorescent labeling; (.) autofluorescence is minimal, even in fixed cells; (.) background staining is min