Protocols for Nucleic Acid Analysis by Nonradioactive Probes: (28 Methods in Molecular Biology)

This book provides an exceptionally useful collection of tried and tested protocols for nucleic acid analysis that avoid the use of radioisotopes. Southern (i.e., DNA) and Northern (i.e., RNA) blotting protocols are covered in detail, from how to isolate good quality nucleic acids from plant and animal sources, to the use of the widest range of probe systems (involving digoxigenin, biotin, fluorescein, horseradish peroxidase labeling, and colorimetric and chemiluminescent detection). Other techniques treated in detail include hybridization in situ to chromosomes and transcribed RNA, newer and less conventional technologies, such as Randomly Amplified Polymorphic DNA (RAPD), reverse dot blots, bacterial identification using magnetic bead and dipstick assays, and nucleic acid sequence-based amplification.

Table of Contents:
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 PolyA+ 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-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. Detection of mRNA in Whole Mounts of Mouse Embryos Using Digoxigenin Riboprobes. Probe Assay-Chemiluminescence Enhanced: A Magnetic Bead Assay for Noncultural Diagnosis of Gonorrhea. 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: The Reverse Dot Blot Method. RAPD Assay: A Novel Technique for Genetic Diagnostics. Nonradioactive Oligonucleotide Probes for Detecting Products of the Ligase Chain Reaction. Nucleic Acid Sequence Based Amplification. Index.

Review :
...this book provides a collection of good protocols which are in general clearly written. They can be easily employed on the bench without being a specialist in a particular fields. - FEBS Letters