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The state-of-the-art methods collected here are designed to analyze the relationship between chromatin structure and function, and to elucidate the molecular mechanisms that control such vital cellular functions as transcription, replication, recombination, and DNA repair. These proven methods cover a wide range of topics, from powerful cell-free systems for the reconstitution of chromatin heterogeneity in vitro, to both classical and cutting-edge techniques for in vivo analysis of protein DNA interactions. Each method includes detailed step-by-step instructions to ensure successful replication along with helpful notes about how to avoid pitfalls. All authors are leading scientists, well known for their methodological expertise in the chromatin research.
Contents
Expression and Purification of Recombinant Histones and Nucleosome Reconstitution Karolin Luger Thomas J. Rechsteiner and Timothy J. Richmond. Preperation and Analysis of Positioned Nucleosomes Vasily M. Studitsky. Site-Directed Chemical Probing of Histone-DNA Interactions David R. Chafin Kyu- Min Lee and Jeff Hayes. Base Pair Resolution Nucleosome Mapping Andrew Flaus and Timothy J. Richmond. Equilibrium and Dynamic Nucleosome Stability Jonathan Widom. Nucleosome Structure and Dynamics: The DNA Minicircle Approach Ariel Prunell Mohamed Alilat Joel Cohen-Solal and Filomena DeLucia. Analysis of Linker Histone Binding to Mono-and Dinucleosomes Simon Chandler and Alan P. Wolffe. Quantitative Analysis of Chromatin Higher Order Organisation Using Agarose Fel Electrophoresis Jeffrey C. Hansen Terace M. Fletcher and Isabelle Kreider. Analytical Ultracentrifugation of Chromatin Jeffrey C. Hansen and Cynthia L. Turgeon. Analysis of Chromatin by Scanning Force Microscopy Sanford Leuba and Carlos Bustamante. In Vivo Mapping of Nucleosomes Using Psoralen-DNA Crosslinking and Primer Extension Ralf E. Wellinger Lucchini Renzo Dammann R. and Jose Sogo. Preparation of Chromatin Assembly Extracts from Xenopus Oocytes David Tremethick. Preparation of Chromatin Assembly Extracts from Preblastoderm Drosphila Embryos Edgar Bonte and Peter B. Becker. A Solid Phase Approach for the Analysis of Reconstituted Chromatin Raphael Sandaltzopoulos and Peter B. Becker. Reconstitution and Analysis of Hyperacetylated Chromatin Wladyslaw A. Krajewski and Peter B. Becker. Assembly of Mitototic Chromosomes in Xenopus Egg Extracts Anne-Elisabeth de la Barem Michel Robert-Nicoud and Stefan Dimitrov. Nucleotide Exicision Repair Coupled To Chromatin Assembly Pierre-Henri Gaillard Daniele Roche and Genevieve Almouzni. Photolyase- A Molecular Tool To Characterise Chromatin Structure in Yeast Fritz Thoma. Transcriptional and Structional Analysis of Isolated SV40 Chromatin Ulla Hansen. In Vitro Replication of Chromatin Templates Claudia Gruss. Analysis of HMG1417- Containing Chromatin Yuri V. Postnikov and Michael Bustin. Identification and Analysis of Native Nucleosomal Histone Acetyltransferase Complexes Patrick A. Grant Shelley Berger and Jerry L. Workman. Analysis of Nucleosome Distribution by ATP-Driven Chromatin Remodeling Complexes Tom Owen-Hughes Rhea T. Utley David J. Steger Joshua M. West Sam John Jacques Cote Kristina M. Havas and Jerry L. Workman. Nucleosome Remodeling Factor NURF and in Vitro Transcription of Chromatin Gaku Mizuguchi and Carl Wu. An SDS-PAGE Based Enzyme Activity Assay for the Detection and Identification of Histone Acetyltranferases James E. Brownell Craig A. Mizzen and C. David Allis. Analysis of DNase I Hypersensitive Sites in Chromatin by Cleavage in Permeabilised Cells Rein Aasland and A Francis Stewart. Mapping of Nucleosome Positions in Yeast Magdalena Livingstone-Zatchej and Fritz Thoma. Analysis of DNA Topology in Yeast Chromatin Randall H. Morse. DNA Methyltransferases as Probes for Chromatin Structure in Yeast Michael P. Kladde and Robert T. Simpson. Restriction Nucleases as Probes for Chromatin Structure Philip Gregory Slobodan Barbaric and Wolfram Horz. Genomic Footprinting Using Nucleases Luca Cappabianca Helen Thomassin Raymond Pictet and Thierry Grange. In Situ Analysis of Chromatin Proteins During Development and Cell Differentiation Usign Flow Cytometry Didier Grunwald Claudia Gorka Sandrine Curtet an Saadi Khochbin. Mapping Target Sites of Chromatin Proteins in Vivo by Formaldehyde Crosslinking Helen Strutt and Renato Paro. Mapping DNA Interactions Sites of Chromosomal Proteins: Crosslinking Studies in Yeast Andreas Hecht Sabine Strahl-Bolsinger and Michael Grunstein. UV Laser Footprinting and Protein-DNA Crosslinking Dimitri Angelov Saadi Khochbin and Stefan Dimitrov. An in Vivo Crosslinking Assay that Detects DNA Binding by Sequence-Specific Transcription Factors Alan Carr and Ma
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