Protocol for the genomic analysis of salt-fractionated chromatin from frozen murine liver

Abstract

Summary: Salt fractionation is a classical approach for separating chromatin based on its differential salt solubility and physical properties. Here, we present a protocol to apply salt fractionation for genome-scale profiling of chromatin isolated from livers at different stages of aging in mice. We elaborate on the steps to isolate nuclei, digest with micrococcal nuclease, sequentially salt fractionate, purify DNA, and construct libraries for genome profiling. We also include information on a computational pipeline for data analysis.For complete details on the use and execution of this protocol, please refer to Yang et al.1 This protocol is an adaptation of the salt fractionation method of Teves and Henikoff.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Genomics
• Molecular Biology