title: GSM8504091: cerebral cortex, Eed-cHet; Mus musculus; RNA-Seq
library_construction_protocol: Tissue was dissected and immediately homogenised in a Dounce homogenizer, filled with 1 ml of homogenisation buffer [250 mM sucrose, 25 mM KCl, 5 mM MgCl2, 10 mM Tris buffer (pH 8), 1 µM DTT, 1x protease inhibitor (cOmplete Protease Inhibitor Cocktail Tablets, Roche), 0.1% (v/v) Triton X-100 10%, and 40 U/mL RNase (RNasIn, Promega), diluted in nuclease-free water]. The homogenate was then incubated in 4 ml of homogenisation buffer for 5 minutes on ice. The homogenate was then strained through a 40 µm cell strainer prewetted with 0.5 ml homogenisation buffer. The strainer was washed with 0.5 ml homogenisation buffer. The homogenate was centrifuged for 5 minutes at 500 g in a swing bucket. The cell pellet was resuspended in 2ml wash media [1% BSA and 100 U/mL RNasIn, diluted in PBS], and DAPI was added at 1:5000 concentration in 1 ml wash media. The cell suspension was submitted for FACS sorting, using DAPI to filter nuclei from debris and to avoid collection of doublet nuclei. The library was prepared with the 10x Genomics Chromium platform using a 3' v3.1 kit. snRNA-Seq