Polycomb Group Proteins: Methods and Protocols

ISBN-13: 9781493981816 / Angielski / Miękka / 2018 / 325 str.

Chiara Lanzuolo; Beatrice Bodega

Spis treści

Mapping the Function of Polycomb Proteins.- Chromatin Immunoprecipitation.- Chromatin preparation and Chromatin Immuno-Precipitation from Drosophila embryos.- ChIP-seq data processing for PcG proteins and associated histone modifications.- Analysis of single locus replication timing in asynchronous cycling cells.- Non coding RNA interplay with the genome.- RIP: RNA Immunoprecipitation.- Capture Hybridization Analysis of DNA Targets.- Identification of RNA-protein interactions through in-vitro RNA pull-down assays.- Understanding RNA-chromatin interactions using Chromatin Isolation by RNA Purification (ChIRP).- Analysis RNA-seq and non coding RNA.- The dynamics of Polycomb complexes.- Fluorescence resonance energy transfer microscopy for measuring chromatin complex structure and dynamics.- Analysis of endogenous protein interactions of Polycomb-group of proteins in mouse embryonic stem cells.- Determination of Polycomb group of proteins compartmentalization through chromatin fractionation procedure.- An Automatic Segmentation Method Combining an Active Contour Model and a Classification Technique for Detecting Polycomb-group Proteins in High-Throughput Microscopy Images.- Polymer physics of the large-scale structure of chromatin.- Chromosome Conformation Capture in Drosophila.- Chromosome Conformation Capture in primary human cells.- Determination of high-resolution 3D chromatin organization using Circular Chromosome Conformation Capture (4C-seq).- Chromosome Conformation Capture on Chip (4C): Data Processing.- In vivo models to address the function of Polycomb Group Proteins.- A rapid TALEN assembly protocol.- Following the motion of Polycomb bodies in living Drosophila embryos.- Reprogramming of Somatic Cells towards Pluripotency by Cell Fusion.- Imaginal discs transplantation in Drosophila.- Isolation and culture of muscle stem cells. 

Blurb

This volume presents the most recent technologies used in the Polycomb Group Proteins (PcG) field.  Chapters detail state-of-the-art methods, creating a unique and comprehensive reference source for investigating Polycomb function in the nucleus. 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, Polycomb Group Proteins: Method and Protocols aims to ensure successful results in the further study of this vital field.