{Reference Type}: Journal Article
{Title}: Protocol to prepare doubly labeled fluorescent nucleosomes for single-molecule fluorescence microscopy.
{Author}: Ghoneim M;Musselman CA;
{Journal}: STAR Protoc
{Volume}: 4
{Issue}: 2
{Year}: 2023 Apr 19
暂无{DOI}: 10.1016/j.xpro.2023.102229
{Abstract}: 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.