Charite - Universitatsmedizin Berlin, Freie Universitat Berlin and Humboldt-Universitat zu Berlin 1 article published in JoVE Biology Nuclei Isolation from Adult Mouse Kidney for Single-Nucleus RNA-Sequencing Janna Leiz*1,2, Christian Hinze*1,2, Anastasiya Boltengagen3, Caroline Braeuning4, Christine Kocks3, Nikolaus Rajewsky3, Kai M. Schmidt-Ott1,2 1Department of Nephrology and Medical Intensive Care Medicine, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, 2Molecular and Translational Kidney Research, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 3Berlin Institute of Medical Systems Biology (BIMSB), Systems Biology of Gene Regulatory Elements, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 4Berlin Institute of Medical Systems Biology (BIMSB), Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC) Here, we present a protocol to isolate high-quality nuclei from frozen mouse kidneys that improve the representation of medullary kidney cell types and avoids the gene expression artifacts from enzymatic tissue dissociation.