GSM8004125: BFP- astrocyte, unsorted, rep1; Homo sapiens; RNA-Seq
Cells were dissociated with Accutase with 10 units/mL of papain (Sigma-Aldrich) for 10 minutes at 37°C and resuspended in 0.5% bovine serum albumin (Sigma Aldrich) with 50 units/mL DNase I in DPBS without calcium or magnesium (Gibco) and passed through a cell strainer with 35 µm mesh. Cells were stained with 1 µM SYTO16 (Invitrogen) and 0.08% (v/v) propidium iodide (part of the Invitrogen ReadyProbes™ Cell Viability Imaging Kit, Blue/Red) for 20 minutes on ice and then dispensed into the nano-well plates of the ICELL8® cx Single-Cell System (Takara). Wells containing single viable cells were automatically selected using the ICELL8 cx CellSelect v2.5 Software (Takara) with the green and not red logic. Manual triage was performed to recover additional candidate wells that contain viable single cells. The cDNA libraries were generated using the SMART-Seq ICELL8 cx Application Kit protocol provided by the manufacturer