DNA Repair Protocols (eBook, PDF)

Redaktion: Henderson, Daryl S.

• Format: PDF

Produktbeschreibung

In this second edition of a much praised laboratory manual devoted to eukaryotic systems, Daryl S. Henderson has refocused the book on mammalian cells, adding fourteen entirely new chapters and extensively revising many of the remaining chapters. The authors address a broad range of questions about practical mammalian DNA repair, including such arcana as "what is radioresistant DNA synthesis and how is it measured?" The techniques presented are readily reproducible and offer cutting-edge methods for cytogenetic analysis, measuring the cellular response to ionizing radiation, detecting single-strand (nicks) and double-strand DNA breaks, detecting the presence of "adducted" bases in DNA, and preparing mismatch repair (MMR) plasmid substrates. Among the highlights are excellent coverage of both base excision repair (BER) and nucleotide excision repair (NER), useful assays for identifying and quantifying UV-induced DNA lesions and DNA breakage, gene therapy, environmental mutagenesis and cancer, and gene targeting. The protocols follow the successful Methods in Molecular Biology(TM) series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Authoritative and highly practical, DNA Repair Protocols: Mammalian Systems, Second Edition, offers investigators a wide variety of productive methods to explore and make new discoveries in the world of mammalian DNA repair.

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Produktdetails

• Verlag: Humana Press
• Seitenzahl: 498
• Erscheinungstermin: 3. Februar 2008
• Englisch
• ISBN-13: 9781592599738
• Artikelnr.: 64701633

Inhaltsangabe

Isolation of Mutagen-Sensitive Chinese Hamster Cell Lines by Replica Plating
Malgorzata Z. Zdzienicka
Complementation Assays Adapted for DNA Repair-Deficient Keratinocytes
Mathilde Fréchet, Valérie Bergoglio, Odile Chevallier-Lagente, Alain Sarasin, and Thierry Magnaldo
Cytogenetic Challenge Assays for Assessment of DNA Repair Capacities
William W. Au and Salama A. Salama
Evaluating the Delayed Effects of Cellular Exposure to Ionizing Radiation
Shruti Nagar, James J. Corcoran, and William F. Morgan
Inhibition of DNA Synthesis by Ionizing Radiation: A Marker for an S-Phase Checkpoint
Nicolaas G. J. Jaspers and Malgorzata Z. Zdzienicka
Analysis of Inhibition of DNA Replication in Irradiated Cells Using the SV40-Based In Vitro Assay of DNA Replication
George Iliakis, Ya Wang, and Hong Yan Wang
Cytometric Assessment of Histone H2AX Phosphorylation: A Reporter of DNA Damage
Xuan Huang and Zbigniew Darzynkiewicz
Detection of DNA Strand Breaks by Flow and Laser Scanning Cytometry in Studies of Apoptosis and Cell Proliferation (DNA Replication)
Zbigniew Darzynkiewicz, Xuan Huang, and Masaki Okafugi
In Vitro Rejoining of Double-Strand Breaks in Genomic DNA
George Iliakis and Nge Cheong
Detection of DNA Double-Strand Breaks and Chromosome Translocations Using Ligation-Mediated PCR and Inverse PCR
Michael J. Villalobos, Christopher J. Betti, and Andrew T. M Vaughan
Plasmid-Based Assays for DNA End-Joining In Vitro
George Iliakis, Bustanur Rosidi, Minli Wang, and Huichen Wang
Use of Gene Targeting to Study Recombination in Mammalian Cell DNA Repair Mutants
Rodney S. Nairn and Gerald M. Adair
Gene-Specific and Mitochondrial Repair of Oxidative DNA Damage
R. Michael Anson, Penelope A. Mason, and Vilhelm A. Bohr
Quantitative PCR-Based Measurement of Nuclear and Mitochondrial DNA Damage and Repair in Mammalian Cells
Janine H. Santos, Joel N. Meyer, Bhaskar S. Mandavilli, and Bennett Van Houten
Measuring the Formation and Repair of DNA Damage by Ligation-Mediated PCR
GerdP. Pfeifer
Immunochemical Detection of UV-Induced DNA Damage and Repair
Marcus S. Cooke and Alistair Robson
A Dot-Blot Immunoassay for Measuring Repair of Ultraviolet Photoproducts
Shirley McCready
Quantification of Photoproducts in Mammalian Cell DNA Using Radioimmunoassay
David L. Mitchell
DNA Damage Quantitation by Alkaline Gel Electrophoresis
Betsy M. Sutherland, Paula V. Bennett, and John C. Sutherland
The Comet Assay: A Sensitive Genotoxicity Test for the Detection of DNA Damage and Repair
Günter Speit and Andreas Hartmann
Fast Micromethod DNA Single-Strand-Break Assay
Heinz C. Schröder, Renato Batel, Heiko Schwertner, Oleksandra Boreiko, and Werner E. G. Müller
32P-Postlabeling DNA Damage Assays: PAGE, TLC, and HPLC
Shinya Shibutani, Sung Yeon Kim, and Naomi Suzuki
Electrophoretic Mobility Shift Assays to Study Protein Binding to Damaged DNA
Vaughn Smider, Byung Joon Hwang, and Gilbert Chu
Construction of MMR Plasmid Substrates and Analysis of MMR Error Correction and Excision
Huixian Wang and John B. Hays
Characterization of Enzymes that Initiate Base Excision Repair at Abasic Sites
Walter A. Deutsch and Vijay Hegde
Base Excision Repair in Mammalian Cells
Yoshihiro Matsumoto
In Vitro Base Excision Repair Assay Using Mammalian Cell Extracts
Guido Frosina, Enrico Cappelli, Monica Ropolo, Paola Fortini, Barbara Pascucci, and Eugenia Dogliotti
Biochemical Assays for the Characterization of DNA Helicases
Robert M. Brosh, Jr. and Sudha Sharma
Repair Synthesis Assay for Nucleotide Excision Repair Activity Using Fractionated Cell Extracts and UV-Damaged Plasmid DNA
Maureen Biggerstaff and Richard D. Wood
Assaying for the Dual Incisions of Nucleotide Excision Repair Using DNA with a Lesion at a Specific Site
Mahmud K. K. Shivji, Jonathan G. Moggs, Isao Kuraoka, and Richard D. Wood
Analysis of Proliferating Cell Nuclear Antigen (PCNA) Associated with DNA Excision Repair Sites in Mammalian Cells
A. Ivana Scovassi

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Mutant Isolation and Gene Cloning.- Isolation of DNA Structure-Dependent Checkpoint Mutants in S. pombe.- Isolating Mutants of the Nematode Caenorhabditis elegans That Are Hypersensitive to DNA-Damaging Agents.- Isolating DNA Repair Mutants of Drosophila melanogaster.- Generation, Identification, and Characterization of Repair-Defective Mutants of Arabidopsis.- Screening for ?-Ray Hypersensitive Mutants of Arabidopsis.- Isolation of Mutagen-Sensitive Chinese Hamster Cell Lines by Replica Plating.- Strategies for Cloning Mammalian DNA Repair Genes.- Novel Complementation Assays for DNA Repair-Deficient Cells.- Recognition and Removal of Inappropriate or Damaged DNA Bases.- The Use of Electrophoretic Mobility Shift Assays to Study DNA Repair.- Mismatch Repair Assay.- Measurement of Activities of Cyclobutane-Pyrimidine-Dimer and (6-4)-Photoproduct Photolyases.- A Dot Blot Immunoassay for UV Photoproducts.- Measurement of UV Radiation-Induced DNA Damage Using Specific Antibodies.- Quantification of Photoproducts in Mammalian Cell DNA Using Radioimmunoassay.- Monitoring Removal of Cyclobutane Pyrimidine Dimers in Arabidopsis.- DNA Damage Quantitation by Alkaline Gel Electrophoresis.- The Comet Assay (Single-Cell Gel Test).- Measuring the Formation and Repair of UV Photoproducts by Ligation-Mediated PCR.- PCR-Based Assays for Strand-Specific Measurement of DNA Damage and Repair I.- PCR-Based Assays for Strand-Specific Measurement of DNA Damage and Repair II.- Gene-Specific and Mitochondrial Repair of Oxidative DNA Damage.- Characterization of DNA Strand Cleavage by Enzymes That Act at Abasic Sites in DNA.- Base Excision Repair Assay Using Xenopus laevis Oocyte Extracts.- In Vitro Base Excision Repair Assay Using Mammalian Cell Extracts.- Nucleotide Excision Repair inSaccharomyces cerevisiae Whole-Cell Extracts.- In Vitro Excision Repair Assay in Schizosaccharomyces pombe.- Nucleotide Excision Repair Assay in Drosophila melanogaster Using Established Cell Lines.- Nucleotide Excision Repair in Nuclear Extracts from Xenopus Oocytes.- Assay for Nucleotide Excision Repair Protein Activity Using Fractionated Cell Extracts and UV-Damaged Plasmid DNA.- Dual-Incision Assays for Nucleotide Excision Repair Using DNA with a Lesion at a Specific Site.- DNA Strand Breakage and Repair.- In Vitro Chemiluminescence Assay to Measure Excision Repair in Cell Extracts.- Physical Monitoring of HO-Induced Homologous Recombination.- Use of P Element Transposons to Study DNA Double-Strand Break Repair in Drosophila melanogaster.- Analyzing Double-Strand Repair Events in Drosophila.- Expression of I-Sce I in Drosophila to Induce DNA Double-Strand Breaks.- Use of I-Sce I to Induce DNA Double-Strand Breaks in Nicotiana.- Chromosomal Double-Strand Breaks Introduced in Mammalian Cells by Expression of I-Sce I Endonuclease.- Induction of DNA Double-Strand Breaks by Electroporation of Restriction Enzymes into Mammalian Cells.- In Vitro Rejoining of Double-Strand Breaks in Genomic DNA.- Extrachromosomal Assay for DNA Double-Strand Break Repair.- Use of Gene Targeting to Study Recombination in Mammalian DNA Repair Mutants.- DNA Damage Tolerance Mechanisms and Regulatory Responses.- Measurement of Low-Frequency DNA Breaks Using Nucleoid Flow Cytometry.- Live Analysis of the Division Cycles in X-Irradiated Drosophila Embryos.- Inhibition of DNA Synthesis by Ionizing Radiation.- Analysis of Inhibition of DNA Replication in Irradiated Cells Using the SV40-Based In Vitro Assay of DNA Replication.- Assays of Bypass Replication of Genotoxic Lesions in Mammalian Disease and Mutant Cell-Free Extracts.- Detection of Chromatin-Bound PCNA in Cultured Cells Following Exposure to DNA-Damaging Agents.- Induction of p53 Protein as a Marker for Ionizing Radiation Exposure In Vivo.- Activation of p53 Protein Function in Response to Cellular Irradiation.- Selective Extraction of Fragmented DNA from Apoptotic Cells for Analysis by Gel Electrophoresis and Identification of Apoptotic Cells by Flow Cytometry.- Detection of DNA Strand Breakage in the Analysis of Apoptosis and Cell Proliferation by Flow and Laser Scanning Cytometry.- Immunoassay for Single-Stranded DNA in Apoptotic Cells.

Rezensionen

"...It will serve as an essential reference for both the practical and theoretical aspects of DNA repair studies, encourage the transfer of methodologies between model systems, and stimulate the development of new approaches." - Anticancer Research"...must for any science library supporting a molecular biology program." - E-Streams"An informative collection of techniques and protocols for characterizing and utilizing DNA repair activities and enzymes from some of the leading names in DNA repair field. This collection of protocols is ideal for both the novice in the field and to the experienced applied researcher." - Lavoisier Librairie

"...an extremely useful resource for any laboratory with an interest in the vibrant field of DNA repair. Any good laboratory manueal should be not only a useful reference guide but also a source of inspiration, and it is undoubtedly true that DNA Repair Protocols: Mammalian Systems succeeds on both counts." -Int. J. Radiation Oncology Biol. Phys.

"This is the definite book on the subject not only for those working in the field of DNA repair but also for those interested in genetic toxicology and chemical safety." -BTS Newsletter

"...a comprehensive series of technique-oriented chapters focusing on eukaryotic DNA repair methodology. ...this text succeeds admirably...The scope of the text is fairly broad and encompasses not only in vitro biochemistry and enzymology, but also cell biology and genetics and even signal transduction....In vitro biochemical assays are well covered,...A particularly nice feature of the book is that the chapters have undergone uniform editing and formatting. All provide a nice overview of the topic, including a brief review of the pertinent literature, followed by step-by-step methods sections. The methods are clearly presented in annotated outline form with cross referencing and high level of detail. Each chapter has a particularly valuable section at the end, called "Notes" in which the authors present some of the nitty-gritty details and tricks of the trade needed to make the techniques work....Overall, this book should provide a valuable laboratory companion for researchers in the area of DNA repair. It serves to provide useful and readable introductions to various topics, along with techniques protocols sufficient for reproducibility....this text will have substantial appeal to the readers of Radiation Research."-Radiation Research

"The list of authors contains many of the leading scientists within the field...Protocols for most experimental eukaryotic organisms are described, from yeast through plants, worms, flies and frogs to mammals. Another laudable quality of this book is the standardization of the descriptions in Materials and methods. Since (almost) all articles are organized similarly, it is relatively easy to find what you want. Also, technical details have been standardized....At the end of each article , there is a 'Notes' section with detailed explanation of specific technical points. For a novice it is good to be reminded that ethidium bromide is a mutagen and that lids should be loosened before putting flasks in the microwaveoven. Iparticularly liked the description, written by the editor, of how to squash Drosophila larvae on a microscope slide by"standing on it with the ball of your foot or your heel. If using the foot method, place the slide (sandwiched in 3 MED MER) on a hard clean floor, cover it carefully with a piece of wood, and stand on that". ...this book keeps up the reputation of the 'Methods in Molecular Biology' series and I would recommend if for labs working with DNA repair, in particular for use by students and technicians."-FEBS Letters