B1 (CRX1805843; CRA027123)
Using single cell 3' Library and Gel Bead Kit V3.1 (10x Genomics, 1000121) and Chromium Single Cell G Chip Kit (10x Genomics, 1000120), the cell suspension was loaded onto the Chromium single cell controller (10x Genomics) to generate single-cell gel beads in the emulsion according to the manufacturer's protocol. Captured cells were lysed and the released RNA were barcoded through reverse transcription in individual GEMs. Reverse transcription was performed on a S1000TM Touch Thermal Cycler (Bio Rad) at 53 degree centigrade for 45 min, followed by 85 degree centigrade for 5 min, and then held at 4 degree centigrade. According to the manufacturer's instructions, single-cell RNA-seq libraries were constructed using Single Cell 3' Library and Gel Bead Kit V3.1