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This volume provides a comprehensive list of protocols for molecular biologists, biochemists and geneticists. Chapters cover protocols that further the study into protein complexes that modify chromatin either by adding or removing post-translational modifications, or by exchanging histone variants within the nucleosome. 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, Histones: Methods and Protocols aims to ensure successful results in the further study of this vital field.
List of contents
In Vitro Assembly of Nucleosomes For Binding/Remodeling Assays.- An Assay for Measuring Histone Variant Exchange within Nucleosomes In Vitro.- Purification of Yeast Native Reagents for the Analysis of Chromatin Function-I: Nucleosomes for Reconstitution and Manipulation of Histone Marks.- Purification of Yeast Native Reagents for the Analysis of Chromatin Function-II: Multiprotein Complexes and Biochemical Assays.- Histone Purification from Saccharomyces Cerevisiae.- Analytical Ultracentrifuge Analysis of Nucleosomes Assembled from Recombinant, Acid Extracted, Hplc-Purified Histones.- Silac-Based Quantitative Strategies for Accurate Histone Post-Translational Modifications Profiling Across Multiple Biological Samples.- Characterization of Individual Histone Post-Translational Modifications and their Combinatorial Patterns by Mass Spectrometry-Based Proteomics Strategies.- Production and Purification of Antibodies against Histone Modifications.- Immunofluorescence of Histone Proteins.- Acid-Urea Gel Electrophoresis and Western Blotting Of Histones.- Chromatin Immunoprecipitation of Histone Modifications in Fission Yeast.- A Spiking Strategy for Chip-Chip Data Normalization in S. Cerevisiae.- High-Resolution Genome-Wide Mapping Of Nucleosome Positioning And Occupancy Level Using Paired-End Sequencing Technology.- Physarum Polycephalum for Studying the Function Of Histone Modifications In Vivo.- A Method for Large Scale Screening of Random Sequence Libraries to Determine the Function of Unstructured Regions from Essential Proteins.
Summary
This volume provides a comprehensive list of protocols for molecular biologists, biochemists and geneticists. Chapters cover protocols that further the study into protein complexes that modify chromatin either by adding or removing post-translational modifications, or by exchanging histone variants within the nucleosome. 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, Histones: Methods and Protocols aims to ensure successful results in the further study of this vital field.
