title: GSM7421574: Cerebellum, replicate 1, snRNA-seq; Sus scrofa domesticus; RNA-Seq
library_construction_protocol: One male and one female Meishan pig, aged 180 days , were obtained from a commercial pig farming company. Each tissue was minced into 5-10 pieces weighing approximately 50-100 mg each on ice with sterilized scissors, which were then snap-frozen in liquid nitrogen and stored at -80℃ until nuclear extraction was performed.Briefly, the tissue samples were homogenized using the Dounce homogenizer with 25 strokes of the loose pestle A and then 25 strokes of the tight pestle B in 1 ml of ice-cold homogenization buffer. After this, the mixture was filtered through a 40-μm cell strainer into a 1.5-ml tube. To collect dissociated single nuclei, the sample was centrifuged at 500g for 5 min at 4℃, and the supernatant was discarded. After centrifugation, the nucleus pellet was resuspended using the desired amount of 1× PBS/0.5% BSA with RNase inhibitor, filtered through a 40-μm cell strainer, and counted. A final concentration of 1,000 nuclei per µl was used for library preparation.