GSM9555374: neuron;CION68; Mus musculus; RNA-Seq
Superficial layers of Sp5C were collected and incubated in a choline chloride solution containing 20 U/mL papain (Worthington) and 100 U/mL DNase I (Worthington) for 15 min at 37°C, and this was followed by digestion with 1 mg/mL protease (Sigma) and dispase (Worthington) for 20 min at 25°C. Next, the post-digested tissues were gently triturated using fire-polished glass Pasteur pipettes to obtain single-cell suspensions. The single-cell suspensions were then centrifuged at 400 g, at 4°C for 5 min, and the supernatant was discarded. The cells were re-suspended with choline chloride solution containing 2% FBS (MACS).Fluorescence labeled cells were manually selected under a microscope and the cytosol of each single cell was transferred individually to a tube. scRNA libraries for scRNA-seq were prepared using Smart-seq3 Protocol V.3 following manufacturer's protocols.(dx.doi.org/10.17504/protocols.io.bcq4ivyw)