GSM5870063: LesionC; Homo sapiens; RNA-Seq
The OLK tissues were washed by phosphate-buffered saline (Gibco, USA), cut into tiny cubes (2mm3), and transferred to the C tube (Miltenyi Biotec, Germany). The tissue dissociation was performed by skin dissociation kits (Miltenyi Biotec, Germany), following the manufacturer's instructions. After dissociation, single-cells were spun down at 400g for 5 min and resuspended with 1X Red Blood Cell Lysis (Abcam, USA) for 5 minutes to remove the blood cells. Then, dead cells removing using Dead Cell Removal kits (STEMCELL Technologies, Canada). Single-cells were spun down at 400g for 5 minutes and adjusted to 1 x 106/ml. The total time from biopsy to loading on the 10X platform was within 3 hours. The cell suspension was loaded into Chromium microfluidic chips with 3' v3, chemistry and barcoded with a 10× Chromium Controller (10X Genomics). RNA from the barcoded cells was subsequently reverse-transcribed and sequencing libraries constructed with reagents from a Chromium Single Cell 3'  v3 reagent kit (10X Genomics) according to the manufacturer's instructions. Sequencing was performed with Illumina NovaSeq according to the manufacturer's instructions (Illumina).