GSM5608517: Hulunbuir Short-tail Sheep embryo; Ovis aries; RNA-Seq
Briefly, tissues were cut into approximately 1-2 mm3 pieces in the RPMI-1640 medium (Gibco), and enzymatically digested with gentleMACS (Miltenyi) using program 37C_m_TDK_1 on a rotor at 37°C, according to manufacturer's instruction. The dissociated cells were subsequently passed through a 100 µm SmartStrainer and centrifuged at 400 g for 5 min. After the supernatant was removed, the pelleted cells were suspended in red blood cell lysis buffer (Miltenyi) and incubated on ice for 1-2 min to lyse red blood cells. After washing twice with 1× PBS (Gibco), the cell pellets were re-suspended in sorting buffer (PBS supplemented with 2% fetal bovine serum (FBS, Gibco)). Cells were loaded around 17,400 cells/chip position using the 10x Chromium Single cell 3' Library, Gel Bead & Multiplex Kit and Chip Kit (10x Genomics, V3.0 barcoding chemistry) according to the manufacturer's instructions.