UID:
almahu_9949251582302882
Format:
1 online resource (XII, 268 p.)
Edition:
1st ed. 1994.
ISBN:
1-59259-515-4
Series Statement:
Methods in Molecular Biology, 28
Content:
In assembling this book, Protocols for Nucleic Acid Analysis by Nonradioactive Probes, I have endeavored to select protocols that have wide applicability. My aim in doing so is to allow nucleic acid analysis to move from the confines of specialized research labora- ries and into general purpose and teaching laboratories. In particular, disciplines such as population biology and clinical diagnostics (where the application of the technology is important, not the technology itself) will find the newer techniques easier to perform with the qu- tity of samples that are normally required in these fields. Die-hard molecular biologists with hot fingers will find, by trying some of the protocols in this book, that nonradioactive protocols are normally faster than their radioactive brethren, and give more reliable results. An expanded description of the subject areas covered by this book is given in Chapter 1. The technology herein has very broad application; for instance, in conventional molecular biology research, population biology, plant and animal breeding, genetic mapping, paternity testing, forensics, prenatal diagnosis, and clinical and food microbiology. I have tried to include comprehensive protocols for both basic and more complex analyses. There are chapters dealing with the fundamentals of nucleic acid extraction from plants and a- mals, and with the procedures necessary to immobilize nucleic acids on solid supports. A range of labeling procedures is included, f- lowed by a selection of hybridization procedures.
Note:
Bibliographic Level Mode of Issuance: Monograph
,
About Nonradioactive Nucleic Acid Detection -- Isolation of DNA from Plants -- Isolation of High-Molecular-Weight DNA from Animal Cells -- Restriction Enzyme Digestion, Gel Electrophoresis, and Vacuum Blotting of DNA to Nylon Membranes -- Isolation of Plant RNA -- Isolation of Total and Poly A+ RNA from Animal Cells -- Preparation of RNA Gel Blots -- Preparation of RNA Dot Blots -- Isolation of Plasmids for the Preparation of Probes -- Production of Hybridization Probes by the PCR Utilizing Digoxigenin-Modified Nucleotides -- Production of DNA Hybridization Probes with Digoxigenin-Modified Nucleotides by Random Hexanucleotide Priming -- Digoxigenin Labeling of RNA Transcripts from Multi- and Single-Locus DNA Minisatellite Probes -- Labeling of Double-Stranded DNA Probes with Biotin -- Preparation of Horseradish Peroxidase-Labeled Probes -- Random Prime Labeling of DNA Probes with Fluorescein-11-dUTP -- Labeling of Oligonucleotides with Fluorescein -- Hybridization of Digoxigenin-Labeled Probes to Southern Blots and Detection by Chemiluminescence -- Southern Blot Hybridization of Digoxigenin-Labeled RNA Minisatellite Probes and Color Detection -- Hybridization and Detection of Digoxigenin Probes on RNA Blots -- Hybridization of Horseradish Peroxidase-Labeled Probes and Detection by Enhanced Chemiluminescence -- Hybridization and Detection of Fluorescein-Labeled DNA Probes Using Enhanced Chemiluminescence -- Hybridization of Fluorescein-Labeled Oligonucleotide Probes and Enhanced Chemiluminescence Detection -- Preparation of Chromosome Spreads by Root-Tip Meristem Dissection for In Situ Hybridization with Biotin-Labeled Probes -- Enzymatic Treatment of Plant Material to Spread Chromosomes for In Situ Hybridization -- Use of Biotin-Labeled Probes on Plant Chromosomes -- Direct Fluorochrome-Labeled DNA Probes for Direct Fluorescent In Situ Hybridization to Chromosomes -- Detection of Digoxigenin-Labeled DNA Probes Hybridized to Plant Chromosomes In Situ -- Preparation of Tissue Sections and Slides for mRNA Hybridization -- Detecting mRNAin Tissue Sections with Digoxigenin-Labeled Probes -- Detection of mRNA in Whole Mounts of Mouse Embryos Using Digoxigenin Riboprobes -- PACE (Probe Assay—Chemiluminescence Enhanced) -- Detection of Foodborne Pathogens Using DNA Probes and a Dipstick Format -- Analysis of Gene Sequences by Hybridization of PCR-Amplified DNA to Covalently Bound Oligonucleotide Probes -- RAPD Assay -- Nonradioactive Oligonucleotide Probes for Detecting Products of the Ligase Chain Reaction -- Nucleic Acid Sequence-Based Amplification (NASBA™).
,
English
Additional Edition:
ISBN 0-89603-254-X
Language:
English
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