Gene

Cell biology

Molecular biology

Biology

Biochemistry

Promoter

Computational biology

Chromatin immunoprecipitation

Chromatin

Micrococcal nuclease

Nuclease

Chemistry

Nucleosome

Immunoprecipitation

Gene expression

eLife

Previously, we described a novel alternative to chromatin immunoprecipitation, CUT&amp;RUN, in which unfixed permeabilized cells are incubated with antibody, followed by binding of a protein A-Micrococcal Nuclease (pA/MNase) fusion protein (Skene and Henikoff, 2017). Here we introduce three enhancements to CUT&amp;RUN: A hybrid protein A-Protein G-MNase construct that expands antibody compatibility and simplifies purification, a modified digestion protocol that inhibits premature release of the nuclease-bound complex, and a calibration strategy based on carry-over of E. coli DNA introduced with the fusion protein. These new features, coupled with the previously described low-cost, high efficiency, high reproducibility and high-throughput capability of CUT&amp;RUN make it the method of choice for routine epigenomic profiling.

Improved CUT&amp;RUN chromatin profiling tools

Improved CUT&amp;RUN chromatin profiling tools

Improved CUT&RUN chromatin profiling tools