Digestion of Oligonucleosomes with Micrococcal Nuclease

Abstract

INI1 is an important protein in the Switch/Sucrose Non-Fermentable (SWI/SNF) complex. It is related to Malignant Rhabdoid tumors (MRT) that appears in infants. Its function is still a mystery, but in children that have MRTs, INI1 is missing or truncated. In this experiment, we digested chromatin from natural cells with Micrococcal nuclease (MNase) so that in future experiments we can run an EMSA to determine if INI1 binds to mono-nucleosomes. Two separate experiments were run with samples containing long oligonucleosome (LON) and different concentrations of MNase. After running titrations of differing concentrations of MNase, it is my conclusion that the proper concentration of MNase to digest chromatin to a single nucleosome is 1×10^-5 units/microliter (units/µL). This concentration produced more mono-nucleosomes and less product of other species.