Laboratory of Genetics and Genomics, National Institute on Aging, NIH, Baltimore, MD 21224, USA.
Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
STAR Protoc. 2024 Jun 21;5(2):103043. doi: 10.1016/j.xpro.2024.103043. Epub 2024 Apr 27.
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. This protocol is an adaptation of the salt fractionation method of Teves and Henikoff..
盐析是一种基于其差异盐溶性和物理性质分离染色质的经典方法。在这里，我们介绍了一种应用盐析方法从不同衰老阶段的小鼠肝脏中分离染色质进行全基因组分析的方案。我们详细说明了分离核、用微球菌核酸酶消化、顺序盐析、纯化 DNA 以及构建基因组分析文库的步骤。我们还包括了数据分析的计算流程信息。有关该方案使用和执行的完整详细信息，请参考 Yang 等人的文章。本方案是对 Teves 和 Henikoff 的盐析方法的改编。
