GSM5396157: human SOD1(G93A) (mutant) 1; Mus musculus; RNA-Seq
Each tissue was finely dissected and incubated in Hibernate A without calcium (BrainBits, Springfield, IL, USA) supplemented with 12 mg papain (Worthington, Lakewood, NJ, USA). This solution is then transferred to a shaker-incubator (set to 30 °C and 60 rpm) for 30 min. After 30 min, using a Pasteur pipette, the solution is triturated three times to release cells from the tissue remnants, and transferred to a new collection tube. The media containing the cells is then gently added to the OptiPrep density gradient (Sigma Aldrich, St. Louis, MO, USA) and centrifuged at 800 g for 15 min at 22 °C. The layers consisting of cells were collected, and this solution was passed through a 40-μm strainer (Fisher Scientific, Hampton, NH, USA). The cells were counted and suspended at a concentration of 100 cells/μl in 0.01% BSA-PBS. Drop-Seq Laboratory Protocol version 3.1