Optimized CUT&RUN protocol for activated primary mouse B cells.

ABSTRACT: ChIP-seq has long been the standard for study of chromatin-protein interactions. However, development of a new technique, CUT&RUN, showed substantial advantages compared to ChIP-seq including higher quality signal while using substantially less sample. While a powerful technique, the original protocol was designed using cell lines and histones as targets. Due to their fragility, this was unsuitable for obtaining high-quality data from activated primary B lymphocytes. To adapt this protocol for B cells, cells were fixed prior to nuclear isolation, and several critical adjustments were introduced to the procedure and reagents. We measured binding of H3K4me3 histone and RNA Polymerase II, detecting robust peaks with as little as 100k nuclei. Additionally, freeze-thaw of B cells prior to proce

SUBMITTER: Ruiz SE 

PROVIDER: S-EPMC12021426 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature