Chromatin Protocols

Comprehensive and up-to-date, this revised second edition is a valuable tool for scientists studying various aspects of chromatin function and an ideal guide to aid in the development of new techniques as well as new ideas in the field of chromatin biology.

Significant advancements have been made in the study of chromatin structure and function over the past fifty years but none as spectacular as those made in the last decade due to the development of novel techniques and the ability to sequence large stretches of DNA. In Chromatin Protocols, Second Edition , expert researchers delineate these cutting-edge techniques via step-by-step laboratory methods and protocols, which encompass a wide array of topics from the isolation of nucleosomes, assembly of nucleosomes and study of the basic chromatin structure to detailed analysis of histone modifications and chromatin function. Written in the highly successful Methods in Molecular Biology™ series style, chapters include brief introductions to the subjects, lists of the necessary materials and reagents, readily reproducible protocols, and Notes sections which highlight tips on troubleshooting and avoiding known pitfalls.

Comprehensive and up-to-date, Chromatin Protocols, Second Edition is a valuable tool for scientists studying various aspects of chromatin function and an ideal guide to aid in the development of new techniques as well as new ideas in the field of chromatin biology.

Provides an easily accessible reference volume for analysis of chromatin structure and function incorporating the latest techniques Includes detailed, step by step guidelines for the analysis of chromatin structure and its role in transcription, replication and other biological processes Details elaborate protocols for the detection and characterization of histone modifications including acetylation, methylation and ubiquitination Describes protocols for techniques ranging from immunohistochemical detection of chromatin modifications to genome-wide analysis of chromatin associated proteins Acts as a comprehensive guide to the most up to date techniques dealing with all aspects of chromatin biology Presents experimental details in an easy to follow modular format typical of the MiMB series Includes supplementary material: sn.pub/extras

Inhalt
Preparation of Chromatin Assembly Extracts from Preblastoderm Drosophila Embryos.- Analysis of Reconstituted Chromatin Using a Solid-Phase Approach.- In Vivo Chromatin Decondensation Assays: Molecular Genetic Analysis of Chromatin Unfolding Characteristics of Selected Proteins.- DNA Methyltransferase Probing of Chromatin Structure Within Populations and on Single Molecules.- Visualization of the Expression of HMGN Nucleosomal Binding Proteins in the Developing Mouse Embryo and in Adult Mouse Tissues.- Drug-Induced Premature Chromosome Condensation (PCC) Protocols: Cytogenetic Approaches in Mitotic Chromosome and Interphase Chromatin.- Analysis of DNA Topology in Yeast Chromatin.- Preparation and Analysis of Uniquely Positioned Mononucleosomes.- Monitoring DNA Breaks in Optically Highlighted Chromatin in Living Cells by Laser Scanning Confocal Microscopy.- Methods to Study Transcription-Coupled Repair in Chromatin.- Cytometric Analysis of DNA Damage: Phosphorylation of Histone H2AX as a Marker of DNA Double-Strand Breaks (DSBs).- Detection of DNA Damage Induced by Topoisomerase II Inhibitors, Gamma Radiation and Crosslinking Agents Using the Comet Assay.- Analysis of Genomic Aberrations Using Comparative Genomic Hybridization of Metaphase Chromosomes.- In Vitro Replication Assay with Mammalian Cell Extracts.- Investigation of Genomic Methylation Status Using Methylation-Specific and Bisulfite Sequencing Polymerase Chain Reaction.- Biochemical Analysis of Arginine Methylation in Transcription.- In Vitro Histone Demethylase Assays.- Histone Chaperone as Coactivator of Chromatin Transcription: Role of Acetylation.- Histone Deacetylase Activity Assay.- In Vitro and In Vivo Assays for Studying Histone Ubiquitination and Deubiquitination.- Detection of Histone H3 Phosphorylation in Cultured Cells and Tissue Sections by Immunostaining.- Chromatin Immunoprecipitation Assays: Analyzing Transcription Factor Binding and Histone Modifications In Vivo.- ChIP on Chip Assays: Genome-Wide Analysis of Transcription Factor Binding and Histone Modifications.- ChIP-on-Chip Analysis Methods for Affymetrix Tiling Arrays.- Non-Radioactive Assay Methods for the Assessment of Telomerase Activity and Telomere Length.- Detecting ATM-Dependent Chromatin Modification in DNA Damage and Heat Shock Response.