FUOAL (CRX2059378; CRA031165)
Single-cell RNA-seq libraries were prepared using the Chromium Single Cell 3' Reagent Kits v3 (10x Genomics) , according to the manufacturer's instructions. Briefly, FACS-sorted cells were washed with 0.04% BSA DPBS for three times and were resuscitated to a concentration of 700~1200 cells/ul (viability >= 85%). Cells were captured in droplets at a targeted cell recovery of cells. After the reverse transcription step, emulsions were broken and Barcoded-cDNA was purified with Dynabeads, followed by PCR amplification. Amplified cDNA was then used for 3'gene expression library construction. For gene expression library construction, 50 ng of amplified cDNA was fragmented and end-repaired, double-size selected with SPRIselect beads, and sequenced on an NovaSeq platform (Illumia) to generate 150 bp paired-end Reads. submitted_instrument_model: Illumina NovaSeq 5000