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Updated: May 9, 2026
Preparation and Fractionation of Xenopus laevis Egg Extracts
Published on: August 27, 2008
Protocol for the purification of replisomes from the Xenopus laevis egg extract system for single-particle cryo-EM
Paolo Passaretti1, Milos A Cvetkovic2, Alessandro Costa2
1Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham, B15 2TT Birmingham, UK.
Abstract:
Here, we present a large-scale FLAG immunoprecipitation protocol to isolate large protein complexes driving DNA replication at replicating chromatin assembled in Xenopus laevis egg extract. We describe how to prepare demembranated sperm nuclei (DNA) and low-speed supernatant egg extract (LSS) and present detailed procedures for sample preparation and application onto grids for negative stain electron microscopy (NS-EM) and cryoelectron microscopy (cryo-EM). For complete details on the use and execution of this protocol, please refer to Cvetkovic et al.1.
