This volume explores the latest technologies used in the Polycomb Group of proteins field and helps scientists--working on PcG proteins--investigate all functions of PcG proteins in diverse cellular contexts. The chapters in this book cover topics such the distribution of histone marks by CUT&Tag in Drosophila embryos; Co-IP in mammalian cells; replication timing of gene loci in different cell cycle phases; STORM and electronmicroscopy and relative data analysis; and polycomb mediated epigenetic modification in spheroids. 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. Cutting-edge and comprehensive, Polycomb Group Proteins: Methods and Protocols, Second Edition is a valuable tool for all researchers looking to expand their knowledge of this developing field.