CiNii Books - The nucleic acid protocols handbook

There can be no doubt that some ofthe most spectacular advances made in science over the past few decades have been in the isolation, analysis, and manipulation of nucleicacids. Thishas ledtoamuchgreaterunderstandingofmechanismsandprocesses across many fields of bioscience, such as biochemistry, microbiology, physiology, pharmacology, and the medical sciences to name a few. It has also led to the growth of the biotechnology industry, which seeks to develop and commercialize many ofthese important processes and methods. Much ofthis has come about because ofthe devel- opment of numerous molecular biology and genetic manipulation techniques. The discovery of restriction enzymes and the development of cloning vectors in the early 1970sopenedthedoortowaysofisolatingandmanipulatingnucleic acidsthathadnever been thought possible. Gene probe labeling and hybridization were developed and refined toprovidepowerfulmethodsofanalysis. These-togetherwiththedevelopment of DNA sequencing methods, protein engineering techniques, and PeR-have all continued to contribute substantially to the understandingofbiological processes at the molecular level. Theprotocols for these importantmethods are the focus ofThe Nucleic AcidProtocols Handbook, whose aim is to provide a comprehensive set oftechniques in onevolume thatwill enable the isolation, analysis, and manipulationofnucleic acids to be readily undertaken. The NucleicAcidProtocols Handbook is divided into 10 parts; within each there are approximately 10chapters. The first fourpartsfollow oneanotherlogically: nucleic acid extraction (Part I), basic separation and analysisofDNA (II), through probe design and labeling (III), and RNA analysis techniques (IV). The following three sections deal with gene libraryconstruction andscreening(V), DNA sequencing (VI), andthe polymerase chain reaction (VII).

Table of Contents

Part I. Nucleic Acid Extraction. Isolation of High-Molecular-Weight DNA from Animal Cells, Ian Garner. Isolation of mRNA by Affinity Chromatography, Sian Bryant and David L. Manning. Isolation and Purification of DNA from Plants, Justin Stacey and Peter G. Isaac. Purification of Uncontaminated, Intact Plant RNA, Shu-Hua Cheng, Brandon D. Moore, and Jeffrey R. Seemann. An Improved Method to Isolate Mitochondrial RNA from Green Plant Tissue, Fei Ye and Ralf Reski. Isolating Chromosomal DNA from Bacteria, Elisabeth Chachaty and Patrick Saulnier. Bacterial DNA Extraction for Polymerase Chain Reaction and Pulsed-Field Gel Electrophoresis, Elisabeth Chachaty and Patrick Saulnier. Isolation of Fungal Nucleic Acids, Surapareddy Sreenivasaprasad. Total RNA Isolation from Bacteria, John Heptinstall. Simultaneous RNA and DNA Extraction from Biopsy Material, Culture Cells, Plants, and Bacteria, Udo Doebbeling. Spectrophotometric Analysis of Nucleic Acids, John Heptinstall and Ralph Rapley. Part II. Basic Separation and Analysis of DNA. Restriction Endonuclease Digestion of DNA, Duncan R. Smith. Agarose Gel Electrophoresis of Nucleic Acids, D. Ross Williams and Ralph Rapley. Preparation of RNA Dot Blots, Rachel Hodge. Native Polyacrylamide Gel Electrophoresis, Adrian J. Harwood. Southern Blotting of Agarose Gels by Capillary Transfer, Ralph Rapley and Jane Davenport -Jones. Pulsed-Field Gel Electrophoresis, John Maule. HPLC of DNA and PCR Products, Elena D. Katz. Part III. Probe Design, Synthesis, and Labeling. End-Labeling of DNA Fragments, Adrian J. Harwood. Nick Translation and Random Hexamer Labeling of DNA, Jane Davenport-Jones. Generation of Labeled Probes by Polymerase Chain Reaction, Y. M. Dennis Lo and Shu F. An. Nonradioactive Oligonucleotide Probe Labeling, Sue Fowler and Ian Durrant. Preparation of Direct, Enzyme-Labeled DNA Probes, Ian Durrant and Timothy Stone. Random Prime Labeling of DNA Probes with Fluorescein-Tagged Nucleotides, Bronwen M. Harvey, Claire B.Wheeler, and Martin W. Cunningham. Hybridization and Detection of Fluorescein-Labeled DNA Probes Using Chemiluminescence, Claire B. Wheeler, Bronwen M. Harvey, and Martin W. Cunningham. Hybridization of Enzyme-Labeled Probes and Detection by Chemiluminescence, Timothy Stone and Ian Durrant. Hybridization and Competition Hybridization of Southern Blots, Rosemary Kelsell. Autoradiography and Fluorography, Eric Quemeneur. Part IV. RNA Analysis Techniques. Formaldehyde Gel Electrophoresis of Total RNA, Sian Bryant and David L. Manning. RNA Probes for the Analysis of Gene Expression, Dominique Belin. Primer Extension Analysis of mRNA, Maggie Walmsley, Mark Leonard, and Roger Patient. S1 Mapping Using Single-Stranded DNA Probes, Stephane Viville and Roberto Mantovani. Measurements of Rate of Transcription in Isolated Nuclei by Nuclear 'Run-Off' Assay, Rai Ajit K. Srivastava and Gustav Schonfeld. One-Tube RT-PCR with Sequence-Specific RT Primers, Ulrich Pfeffer and Paola Ferro. Characterization of RNA Using Continuous RT-PCR Coupled with ELOSA, Francois Mallet. Quantitative Analysis of RNA Species by Polymerase Chain Reaction and Solid-Phase Minisequencing, Anu Suomalainen and Ann-Christine Syvanen. Nonradioactive Northern Blotting of RNA, Rainer Loew. Analysis of RNA by Northern Blotting Using Riboprobes, Rai Ajit K. Srivastava. Part V. Gene Library Construction and Screening. Production of Double-Stranded cDNA for Gene Library Synthesis, Jane Kirk and Steve Mayall. Using Rapid Amplification of cDNA Ends (RACE) to Obtain Full-Length cDNAs, Yue Zhang and Michael A. Frohman. cDNA Library Construction Using Streptavidin-Paramagnetic Beads and PCR, Kris N. Lambert and Valerie M. Williamson. Rapid (Ligase-Free) Subcloning of Polymerase Chain Reaction Products, Alan R. Shuldiner and Keith Tanner. Subtraction Hybridization cDNA Libraries, Clifford W. Schweinfest, Peter S. Nelson, Michael W. Graber, Rita I. Demopoulos, and Takis S. Papas. Cloning Polymerase Chain Reaction

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The nucleic acid protocols handbook

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