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This book details recently developed technologies and conventionally employed cytological proceduresfor the study of X-Chromosome Inactivation. Chapters detail live imaging, bioinformatic methods, fluorescence in situ hybridization, and immunofluorescence, and procedures to optimize the study of molecular mechanism underlying X chromosome inactivation. 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, X-Chromosome Inactivation: Methods Protocols aims to be useful for researchers in the field of epigenetics, chromatin, noncoding RNA, and nuclear architecture.
List of contents
Screening for Factors Involved in X Chromosome Inactivation using Haploid ESCs.- Unbiased Genetic Screen to Identify Factors Involved in X-chromosome Inactivation Using a Pooled Bar-coded shRNA Library.- Unbiased Genetic Screen to Identify Factors Involved in X-chromosome Inactivation Using a Pooled Bar-coded shRNA Library.- Manipulation of Xist Imprinting in Mouse Pre-implantation Embryos.- Somatic Cell Nuclear Transfer in Mice - Basic Protocol and Its Modification for Correcting X Chromosome Inactivation Status.- Live Imaging of Xist RNA.- Live Imaging of X-chromosome Inactivation and Reactivation Kinetics.- Visualizing the Dynamics of Inactive X chromosomes in Living Cells Using Antibody-based Fluorescent Probes.- Visualization of Hidden Epitopes at the Inactive X Chromosome.- Chromosome Spread Analyses of Meiotic Sex Chromosome Inactivation.- Simultaneous RNA-DNA FISH in Mouse Pre-implantation Embryos.- Combined Immunofluorescence, RNA FISH and DNA FISH in Pre-implantation Mouse Embryos.- RNA-FISH and Immunofluorescence of Mouse Pre- and Post-implantation Embryos.- Experimental Analysis of Imprinted Mouse X-chromosome Inactivation.- X-chromosome Inactivation and Escape from X inactivation in Mouse.- Practical Analysis of Hi-C Data: Generating A/B Compartment Profiles.
