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丁香实验推荐阅读
Detection of HCV RNA in Formalin-Fixed, Paraffin-Embedded Liver Tissue by RT-PCR

Hepatitis C virus (HCV) RNA has been detected in the sera and liver sections of patients with chronic HCV infection using RT-PCR and other sensitive molecular techniques. Since archived formalin-fixed, paraffin embedded tissues from these patients are readily available for retrospe ...

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Quantification of HCV RNA in Liver Tissue by bDNA Assay

With this statement, Sherlock and Dooley have described two of the three major challenges involved in quantitatively measuring any analyte in tissue samples: the distribution of the analyte in the tissue; and the standard of reference, or denominator, with which to make comparisons betwe ...

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Preparation of Genotype-Specific HCV RNA Transcripts for Assessing HCV Detection and Quantification Assays

Hepatitis C virus (HCV), the etiological agent responsible for the majority of cases of parenterally acquired liver disease, is found throughout the world. HCV is an enveloped virus with a small, single-stranded RNA genome. Because it uses an error-prone, RNA-dependent RNA polymerase, HCV h ...

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Hepatitis C Virus: Types, Subtypes, and Beyond

Non-A, non-B hepatitis was recognized as a frequent consequence of blood transfusion for many years before the agent responsible, hepatitis C virus (HCV), was first cloned and sequenced in 1989 Very quickly it became apparent that viruses from different parts of the world were distinct, and aft ...

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Molecular Evolutionary Analysis: Its Application in the Study of Hepatitis C Virus

The recent development of high-power personal computer hardware and software has allowed investigators to prepare phylogenetic trees without understanding of the fundamental principles of molecular evolution. Unfortunately, inexperienced investigators are also pr ...

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Genotyping by Type-Specific Primers That Can Type HCV Types 16

Hepatitis C virus (HCV) was identified as a major causative agent of non-A, non-B hepatitis (1). Numerous complete or partial nucleotide sequences of HCV isolates have been reported worldwide, and comparison of these sequences revealed their marked genetic heterogeneity nature, sugge ...

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Genotyping Hepatitis C Virus by Type-Specific Primers for PCR Based on NS5 Region

Hepatitis C virus (HCV), a flavi-like virus with a positive-sense, single-stranded genome RNA (1–3), shows considerable variation in nucleotide sequences. These variations fall into a series of specific patterns and are the basis for classification of HCV into different types and genoty ...

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Determination of HCV Genotypes by RFLP

Several different methods have been developed for the typing of HCV variants: direct sequence analysis, slot-blot hybridization analysis of reverse transcriptase-polymerase chain reaction (RT-PCR) products using cDNA probes specific to each HCV genotype and PCR amplificati ...

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HCV Genotyping by the Line Probe Assay INNO-LiPA HCV II

Hepatitis C viruses (HCVs) constitute a highly variable genus within the Flaviviridae, with closest homology to the hepatitis G and GB viruses, and Pestiviruses. The positive-stranded RNA genome encodes a polyprotein which is co- and posttranslationally cleaved into at least nine prot ...

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Serological Genotyping Using Synthetic Peptides Derived from the NS4 Region

The RNA genome of hepatitis C virus (HCV) displays extensive sequence variation, and consequently, the virus is classified into six major genotypes. The severity of disease and response to antiviral treatment are thought to be influenced by both viral and host-related factors, including a ...

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Determination of HCV Quasispecies by Cloning and Sequencing

Most RNA viruses exists as a heterogeneous mixture of closely related viral genome in the host, which is the result of high error rates in RNA replication and selected by various viral and host factors. The spectrum of this spectrum of related genomes within a host is referred as quasispecies (1). This im ...

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Hepatitis C Virus Heteroduplex Tracking Assay: Application to Genotype Determination, Quasispecies Analysis, and Molecular Evolution Studies

The heteroduplex tracking assay (HTA) is a tool that can be used for determining genotype, quasispecies analysis, molecular evolution, and epidemiological studies (1–7). By hybridizing a labeled, single-stranded DNA probe to colinear, reverse transcriptase (RT) PCR products from a s ...

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Detection of Hepatitis C Virus Quasispecies Heterogeneity by Single-Strand Conformational Polymorphism

Hepatitis C virus (HCV) is believed to replicate via a viral-encoded, RNA-dependent RNA polymerase. This replication strategy has limited fidelity Thus, HCV is genetically heterogenous. To date, six HCV genotypes and more than 80 viral subtypes have been identified. Further, even within i ...

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In Situ Detection of HCV: An Overview

Hepatitis C virus (HCV) is the agent responsible for the majority of cases of the parenterally transmitted non-A, non-B hepatitis. The major obstacles to its discovery were the low level of replication in the infected host, both natural and experimental, and the low immunogenicity of its protei ...

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In Situ Detection of Hepatitis C Viral Antigens

A reliable detection system for HCV antigens in liver tissue may be used to identify the HCV cellular tropism and subcellular sites of viral replication Also, it can be used to study the relationship between viral expression and disease activity. Finally, it can facilitate the study of host-viral ...

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In Situ Hybridization and the Detection and Localization of HCV RNA

The procedure described below was originally reported to detect the hepatitis C virus RNA (genomic strand) by nonradioisotopic in situ hybridization in formalin-fixed, paraffin-embedded liver tissue of two acutely infected chimpanzees, in a collaborative study conducted with R. ...

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High-Purity Plasmid Isolation Using Silica Oxide

The isolation of plasmid DNA from bacteria is a crucial technique in molecular biology and is an essential step in many procedures such as cloning, DNA sequencing, transfection, and gene therapy. These manipulations require the isolation of high-purity plasmid DNA. Commercial anion-ex ...

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High-Throughput Plasmid Extraction Using Microtiter Plates

Plasmid extraction is typically performed to produce template DNA for a desired molecular biological reaction, or set of reactions, such as restriction endonuclease digestion (see Chapter 20), DNA sequencing (see Chapter 22), in vitro mutagenesis (see Chapters 23–26), transformat ...

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Isolation of Cosmid and BAC DNA from E. coli

Cosmid and bacterial artificial chromosome (BAC) systems have been developed for the cloning of large DNA inserts averaging 40 kb and 130 kb (range: 90–300 kb), respectively. The resulting clones are more stable than yeast artificial chromosomes (YACs) and rarely chimeric, which makes them e ...

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Preparation of Single-Stranded DNA from Phagemid Vectors

Single-stranded DNA (ssDNA) is the optimal template for most polymerase-based molecular-biology applications, including DNA sequencing and site-directed mutagenesis. Phagemids are chimeric vectors, derived from the ssDNA bacteriophages M13, fd, or f1, that normally repli ...

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