GSM8489345: Frontal cortex, P7-P9, replicate 6; Mus musculus; RNA-Seq
Brains were removed and placed in Hibernate media on ice. FC and VC were dissected, cut into 0.5mm pieces and transferred to fresh HABG at 37oC for 10 min. The tissue was then incubated in papain in Hibernate A without calcium for 30 min at 30C. The papain was replaced with 2 ml of HABG media, and cells were dissociated by trituration. The trituration step was repeated twice and the final cell suspension was passed through a Falcon 40-micron cell strainer, followed by centrifugation at 200g for 10 min. The cells were resuspended in HABG to yield a final concentration of 800-1000 cells/mL and >80% viability (trypan blue exclusion). Library preparation was performed in the Spatial Biology and Genomics Core at the Saban Research Institute, CHLA. Each replicate was processed through Chromium Next GEM Automated Single Cell 3′ Library and Gel Bead Kit v3.1, using the manufacturer's protocol, with an output target of 10,000 cells.