Main description:

This volume presents a comprehensive collection of quick assays for the detection of nuclear and mitochondrial DNA damage and its effects in live and fixed cells and tissues, and in bacterial genomes. Although, such rapid techniques are in demand in the "research trenches" they are not covered well in the literature. This volume is the first such compendium of the time-saving techniques for detection of DNA damage and its direct physiological outcomes including apoptosis, necrosis and phagocytic clearance. The volume demonstrates all levels of detection, starting from the molecular level up to the level of the entire live organism. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Fast Detection of DNA Damage: Methods and Protocols aims to provide easily reproducible techniques requiring only few steps to perform.

Contents:

Part I: EXPRESS DETECTION (from 3 minutes to 1 hour) 1. Express FRET Labeling and Analysis of Phagocytic Clearance

Vladimir V. Didenko

2. Rapid Assessment of Genotoxicity by Flow Cytometric Detection of Cell Cycle Alterations

Nevenka Bihari

3. Ultrasound Imaging of DNA-Damage Effects in Live Cultured Cells and in Brain Tissue Hadi Tadayyon, Mehrdad J. Gangeh, Roxana Vlad, Michael C. Kolios, and Gregory J. Czarnota

4. Ultrasound Imaging of Apoptosis: Spectroscopic Detection of DNA-Damage Effects In Vivo

Hadi Tadayyon, Mehrdad J. Gangeh, Roxana Vlad, Michael C. Kolios, and Gregory J. Czarnota

5. Fluorochrome-Labeled Inhibitors of Caspases: Expedient In Vitro and In Vivo Markers of Apoptotic Cells for Rapid Cytometric Analysis

Zbigniew Darzynkiewicz, Hong Zhao, H. Dorota Halicka, Piotr Pozarowski, and Brian Lee

6. The Fast-halo Assay for the Detection of DNA Aamage

Piero Sestili, Cinzia Calcabrini, Anna Rita Diaz, Carmela Fimognari, and Vilberto Stocchi

7. Rapid Detection of Bacterial Susceptibility or Resistance to Quinolones

Fatima Otero, Rebeca Santiso, Maria Tamayo, German Bou, Jaime Gosalvez, and Jose Luis Fernandez

8. Rapid Detection of Apoptosis in Cultured Mammalian Cells

Kudryavtsev I., Serebryakova M., Solovjeva L., Svetlova M., and Firsanov D

9 Quick Detection of DNase II-type Breaks in Formalin-Fixed Tissue Sections

Candace. L. Minchew and Vladimir V. Didenko

Part II: RAPID DETECTION (3 hours or less)

10. Express -H2AX Immunocytochemical Detection of DNA Damage

Nate Hopp, Jodi Hagen, Birte Aggeler, and Alexander E. Kalyuzhny

11. Rapid detection of -H2AX by Flow Cytometry in Cultured Mammalian Cells Firsanov D., Solovjeva L., Lublinskaya O., Zenin V., Kudryavtsev I., Serebryakova M., and Svetlova M.

12. Rapid Detection of DNA Strand Breaks in Apoptotic Cells by Flow- and Image- Cytometry

Hong Zhao and Zbigniew Darzynkiewicz

13. Fast Micromethod: Determination of DNA Integrity in Cell Suspensions and in Solid Tissues

Nevenka Bihari

14. Quantification of DNA Damage and Repair in Mitochondrial, Nuclear, and Bacterial Genomes by Real-Time PCR

Nadja Patenge

15. Zebra Tail Amplification: Accelerated Detection of Apoptotic Blunt Ended DNA Breaks by In Situ Ligation

Vladimir V. Didenko

Part III: ACCELERATED DETECTION (time-saving versions of conventional techniques)

16. Twelve Gel Comet Assay Format for Quick Examination of DNA Damage and Repair

Sergey Shaposhnikov and Andrew Collins

17. Immunofluorescence Analysis of -H2AX Foci in Mammalian Fibroblasts at Different Phases of the Cell Cycle

Solovjeva L., Firsanov D., Pleskach N., and Svetlova M.

18. RAPD-PCR as a Rapid Approach for the Evaluation of Genotoxin-Induced Damage to Bacterial DNA

Rosanna Tofalo and Aldo Corsetti

19. Rapid Detection of -H2Av Foci in ex vivo MMS-treated Drosophila Imaginal Discs

Varandt Y. Khodaverdian and Mitch McVey