RNA-Seq of Homo sapiens: adult male benign prostatic hyperplasia single cells: Sample 201-BPH-FACS
"Fresh prostate tissue was acquired from radical prostatectomy cases, where BPH was identifiable grossly by transition zone nodules, and normal prostate tissue harvested from the peripheral zone opposite that having biopsy-confirmed cancer. Tissue was disaggregated using collagenase/hyaluronidase, trypsin and dispase, following StemCell Technologies Prostate Tissue Dissociation protocol. Contaminating red blood cells were lysed using ammonium chloride solution (StemCell Technologies), and single cells filtered through a 40uM cell strainer. Half the remaining cells were further purified by flow cytometry, gating on viable (propidium iodide-negative) single cells. Single-cell RNAseq was then performed using the 10X Genomics Chromium workflow (by the Stanford Functional Genomics Facility), entailing microdroplet single cell capture (with cell and molecular barcodes), cDNA synthesis, pooling and library preparation, followed by sequencing on an Illumina HiSeq4000."
alignment_software
"Cell Ranger, version 2"