GSM6890194: Pca_01_N; Homo sapiens; RNA-Seq
Tissue biopsies were extracted within one hour after surgery from prostate specimens. The biopsies were rinsed with PBS, minced into 1mm3 pieces and transferred to the 5 ml of digestion solution ((DMEM high glucose (Sigma-Aldrich) supplemented with with L-glutamine-penicillin-streptomycin (100 U/ml) (Sigma-Aldrich), 20 uL DNase (Quiagen) (75 U/ml), 2 mL Collagenase V (2 %) (Sigma-Aldrich) and Dispase II (2.5 U/mL) (Gibco)) for 45 minutes in water bath (37 °C). To perform scRNA sequencing, 10X Chromium Chip Single-cell library generation and preparation were performed on the single-cell suspension according to 10X Chromium 3'solution (V2 kit) as per manufacturer's instructions with an aim to capture 5000-10000 cells/channel. Sequencing was performed using the Illumina HiSeq 4000 platform. Raw sequencing data was processed by Cell Ranger software (10X Genomics, version 3.1) and aligned to the human reference genome (GRCh38).