Isolation of Pure Nuclei Using a Sucrose Method

This protocol describes a modified version of a widely used method to isolate nuclei from tissue culture cells. It involves mechanical homogenization of cells in isotonic sucrose, followed by velocity centrifugation of nuclei through a denser layer of sucrose. This method, which yields highly pure and intact nuclei, can be optimized for use in various types of tissues and cells. Limitations of the method, alternative options for homogenization, and recommendations for the use of detergents are discussed.