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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.

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1. In Vitro Assembly of Nucleosomes For Binding/Remodeling Assays

Arnob Dutta and Jerry L. Workman

2. An Assay for Measuring Histone Variant Exchange within Nucleosomes In Vitro

Liette Laflamme, Benoit Guillemette, and Luc Gaudreau

3. Purification of Yeast Native Reagents for the Analysis of Chromatin Function-I: Nucleosomes for Reconstitution and Manipulation of Histone Marks

Nicolas Lacoste, Wajid Bhat, and Jacques Côté

4. Purification of Yeast Native Reagents for the Analysis of Chromatin Function-II: Multiprotein Complexes and Biochemical Assays

Nicolas Lacoste, Wajid Bhat, and Jacques Côté

5. Histone Purification from Saccharomyces Cerevisiae

Frederic Jourquin and Vincent Géli

6. Analytical Ultracentrifuge Analysis of Nucleosomes Assembled from Recombinant, Acid Extracted, HPLC-Purified Histones

Manjinder S. Cheema and Juan Ausió

7. Silac-Based Quantitative Strategies for Accurate Histone Post-Translational Modifications Profiling Across Multiple Biological Samples

Alessandro Cuomo, Monica Soldi , and Tiziana Bonaldi

8. Characterization of Individual Histone Post-Translational Modifications and their Combinatorial Patterns by Mass Spectrometry-Based Proteomics Strategies

Simone Sidoli and Benjamin A. Garcia

9. Production and Purification of Antibodies against Histone Modifications

Benoit Guillemette, Ian Hammond-Martel, Hugo Wurtele, and Alain Verreault
10. Immunofluorescence of Histone Proteins
Hilmar Strickfaden and Michael J. Hendzel

11. Acid-Urea Gel Electrophoresis and Western Blotting Of Histones

Catherine A. Hazzalin and Louis C. Mahadevan

12. Chromatin Immunoprecipitation of Histone Modifications in Fission Yeast

Jean Mbogning and Jason C. Tanny

13. A Spiking Strategy for Chip-Chip Data Normalization in S. Cerevisiae

Célia Jeronimo and Fran‡ois Robert

14. High-Resolution Genome-Wide Mapping Of Nucleosome Positioning And Occupancy Level Using Paired-End Sequencing Technology

Mylène Brunelle, Sébastien Rodrigue, Pierre-Étienne Jacques, and Nicolas Gévry

15. Physarum Polycephalum for Studying the Function of Histone Modifications In Vivo

Vanessa Menil-Philippot and Christophe Thiriet
16. A Method for Large Scale Screening of Random Sequence Libraries to Determine the Function of Unstructured Regions from Essential Proteins
Jean-Fran‡ois Millau, Benoit Guillemette, and Luc Gaudreau

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