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This collection of readily reproducible techniques for repairing mammalian DNA contains fourteen entirely new chapters and many extensively revised chapters. The methods presented cover 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.
Isolation of Mutagen-Sensitive Chinese Hamster Cell Lines by Replica Plating; Malgorzata Z. Zdzienicka; Complementation Assays Adapted for DNA Repair-Deficient Keratinocytes; Mathilde Frechet, Valerie Bergoglio, Odile Chevallier-Lagente, Alain Sarasin, and Thierry Magnaldo; Cytogenetic Challenge Assays for Assessment of DNA Repair Capacities; William 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 Ma_gorzata 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 Chromosomal Translocations Using Ligation-Mediated 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 DNA Repair Mutants; Rodney Nairn and Gerald 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; Gerd P. Pfeifer; Immunochemical Detection of UV-Induced DNA Damage and Repair; Marcus S. Cooke and Alistair Robson; A Dot-Blot Immunoassay for Measuring Repair of UV 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; Gunter Speit and Andreas Hartmann; Fast Micromethod DNA Single-Strand Break Assay; Heinz C. Schroder, Renato Batel, Heiko Schwertner, Oleksandra Boreiko, and Werner E. G. Muller; 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; Analysis of Enzymes That Act At Abasic Sites in DNA; 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, Barbar 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 and Ennio Prosperi; Analysis of DNA Repair and Chromatin Assembly In Vitro Using Immobilized Damaged DNA Substrates.
...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