Multiplexed scNOMe-seq protocol based on isolated single nuclei v2

Abstract

What follows is the protocol for performing single-cell Nucleosome Occupancy and Methylome sequencing on single nuclei (scNOMe-seq). This plate-based method, which can be used to simultaneously assess nucleosome occupancy and DNA methylation in single nuclei, draws upon the approach laid out in Luo et al., 2018. Everything from the initial nuclei sorting step to the final library preparation that directly precedes sequencing is included in this protocol. This protocol is optimized for non-neuronal nuclei and can be applied to a range of different cell types (e.g., intestinal organoid, fibroblasts). This is the method we are using and it is working.