Nucleic Acid Probes and Hybridization, Molecular Hybridization Techniques, Immuological Techniques, Extrachromosomal Nucleic Acid

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Nucleic Acid Probes & Hybridization Nucleic Acid Probes & Hybridization Introduction Nucleic Acid Probes commonly Used Radioisotops with Their Half Lifes Probe Development Choice of Probes DNA Probes RNA Probes Oligonucleotides Probes Labelling of Probes Radioactive Probes Labelling of DNA Random Priming Labelling of DNA by Nick Translation End Labelling at 5' End End Labelling at 3' End Non Radioactive Probes Removal of Unincorporated Nucleotides Uses of Nucleic Acid Probes Detection of Pathogens in Clinical Samples Differentiation of Virulent from Avirulent Organisms Typing of Microorganisms Mapping Genes Epidemilogical Studies Food Safety Detection of Contaminant in Culture Study of Mechanisms of Pathogenesis Genetic Analysis Hybridization Hybridization Strategies Factors Affecting the Rate of Hybridization Nitrocellulose Filters - NC Uncharges Nylon Filters - NF Propeprties of Nylon, Nitrocellulose and Charged Nylon Filters Charged Nylon Filters

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	Introdcution Advances in recombinant DNA technology have directly led to the development of widespread application of molecular hybridization techniques in diagnostic and molecular biology laboratories. A wide variety of microbial infections as well as genetic disorders are now diagnosed using hybridization techniques (Haase et al., 1984; Mulchay, 1986, Mc Dougall et al., 1986; Caskay, 1987). In molecular hybridization, a specific nucleic acid probe is used to hybridize to complementary nucleic acid of a specific pathogen species directly in specimens or to analytes immobilized on solid phases. For genotype analysis, cloned genomic sequences are used to detect restriction fragment length polymorphisms in Southern blot hybridization protocols. Probes are labelled with either radionucleotide or biotin in order to permit detection of specific hybrids.
Hybridization is particularly useful in the diagnosis of microbial infections in which the pathogen cannot be identified using conventional or classical diagnostic procedures. Some pathogens cannot be propagated in laboratory bioassays. Some viral pathogens may be present in latent states or integrated into host genomes. In such circumstances, transcriptional and translational processes are typically minimal and infectious agents may not be recoverable.
Two techniques are now widely used to diagnose infections by directly identifying the pathogen in clinical specimens: antigen detection using immunological techniques and nucleic acid detection using hybridization. Although both methods have proven to be sensitive and reliable for providing diagnostic results, molecular hybridization does permit diagnosis of certain infections and conditions where antigen detection cannot. For example, integrated or extrachromosomal nucleic acid of pathogens can be detected by hybridization, permitting diagnosis in the absence of gene expression.

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Immobilization of Nucleic Acid on Filters Immobilization by Baking Immobilization by UV Fixation Immobilization by Alkali Fixation Prehybridization, Hybridization and Washing Detection of Radioactive Hybrids Detection of Non Radioactive Hybrids Chromogenic Substrates Chemiluminescent Substrates Enhanced Chemiluminescence - ECL Solution Hybridization Filter Hybridization Probing Recombinant libraries Southern Blots Northern Blots Dot / Slot Blots In Situ Hybridization DNA Arrays and Chips Preapration of DNA Arrays Preparation of Oligonucleotide Arrays Applications of DNA Arrays Detection of Mutation and Polymorphisms DNA Sequencing by Hybridization Microarrays for Diagnosis Hybridization in Diagnosis Diagnosis of Bovine Herpesvirus Infection Using Biotinylated cDNA Probes Diagnosis of Enterotoxigenic E.Coli Infection Diagnosis of Sickle Cell Anaemia DNA Finger Printing for Genotype Analysis

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