%0 Journal Article
%T Protocol to prepare doubly labeled fluorescent nucleosomes for single-molecule fluorescence microscopy.
%A Ghoneim M
%A Musselman CA
%J STAR Protoc
%V 4
%N 2
%D 2023 Apr 19
%M 37083320
暂无%R 10.1016/j.xpro.2023.102229
%X Single-molecule fluorescence microscopy (SMFM) has been shown to be informative in understanding the interaction of chromatin-associated factors with nucleosomes, the basic building unit of chromatin. Here, we present a protocol for preparing doubly labeled fluorescent nucleosomes for SMFM. We describe steps for over-expression in E. coli and purification of recombinant human core histones. We then detail fluorescent labeling of histones and nucleosomal double-stranded DNA followed by octamer refolding and nucleosome reconstitution.