Full Description
The volume outlines techniques used to study the primary mechanisms of important DNA repair pathways. The chapters in this book are organized into five sections, each discussing a specific aspect of repair biology. Part I reviews the role of post-translational modifications of HDR, and DNA damage caused by defective telomere replication. Part II describes methods to detect and measure DNA breaks and proteins involved in DNA damage response. Part III talks about methods designed to measure DNA repair efficiency and characterize factors involved in the repair process. Part IV discusses various in vitro assays used to purify recombinant proteins that are used to study DNA repair enzyme mechanisms. Lastly, Part V focuses on methods to study the damage arising during DNA replication. 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.
Cutting-edge and authoritative, DNA Repair: Methods and Protocols is an essential resource for novice and experienced researchers, and provides easily adaptable techniques to help them advance in their specific protein or repair pathway of interest.
Contents
Homologous Recombination-Mediated DNA Repair and Implications for Clinical Treatment of Repair Defective Cancers.- Resolving Roadblocks to Telomere Replication.- In Time and Space: Laser Micro-Irradiation and the DNA Damage Response.- Quantification of Double Strand Breaks in Mammalian Cells using Pulsed-Field Electrophoresis.- Methods to Study Trinucleotide Repeat Instability Induced by DNA Damage and Repair.- Extracting and Measuring dNTP Pools in Saccharomyces cerevisiae.- Conversion Tract Analysis of Homology-Directed Genome Editing using Oligonucleotide Donors.- Reporter Assays for BER Pathways.- Methods for Studying DNA Single-Strand Break Repair and Signaling in Xenopus laevis Egg Extracts.- Chromatin Immunoprecipitation (ChIP) of Plasmid-Bound Proteins in Xenopus Egg Extracts.- Cellular Assays to Study the Functional Importance of Human DNA Repair Helicases.- A Mammalian Genetic Complementation Assay for Assessing Cellular Resistance to Genotoxic Compounds.- Small Molecule Inhibitor Screen for DNA Repair Proteins.- Assembling the Human Resectosome on DNA Curtains.- Thin-Layer Chromatography and Real-Time Coupled Assays to Measure ATP Hydrolysis.- Gel-Based Assays for Measuring DNA Unwinding, Annealing, and Strand Exchange.- In Vitro Assay for Plasmid Length DNA Strand Exchange by Human DMC1.- In Vitro Assays for DNA Branch Migration.- Stabilization of Human DMC1 Nucleoprotein Filament.- Measuring UV Photoproduct Repair in Isolated Telomeres and Bulk Genomic DNA.- Single-Molecule DNA Fiber Analyses to Characterize Replication Fork Dynamics in Living Cells.- Direct Visualization of DNA Replication at Telomeres using DNA Fiber Combing Combined with Telomere FISH.- Telomere and G-Quadruplex Co-Localization Analysis by Immunofluorescence-Fluorescence In Situ Hybridization (IF-FISH).- FISHing for Damage on Metaphase Chromosomes.
