Protocol for enhancing RNA yield and quality from single nuclei isolated from mouse brain tissue

Isolating RNA from single nuclei is essential for single-cell gene expression analysis, yet obtaining high-quality RNA is challenging. We present a protocol to enhance RNA yield and quality from mouse brain nuclei. Key steps include brain dissection, thawing, homogenization, and centrifugation-based isolation. The protocol incorporates 3% glyoxal fixation for RNA preservation, followed by filtration, blocking, and fluorescence-activated sorting to ensure the extracted RNA meets quality and quantity standards for transcriptomic and qPCR analyses. For complete details on the use and execution of this protocol, please refer to Marin-Valencia, Kocabas et al.¹