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The Singulator™ 100 can be used to quickly and efficiently isolate nuclei from a wide variety of tissue types, and from samples that are fresh, frozen, or OCT preserved. If the nuclei are intended for downstream RNA analysis, it is critical that care be taken to prevent degradation of the RNA in the nuclei by the activity of endogenous RNase enzymes, most importantly RNase A. To help preserve RNA integrity, RNase inhibitor is commonly added to samples during the isolation and purification of nuclei. Here we describe the use of RNase inhibitors with the Singulator for nuclei isolations.