GSM8774589: Epithelial cells, Pten KO1; Mus musculus; RNA-Seq
For RNA-seq, epithelial cells were stained with EPCAM, viability dye, and 7-AAD, and viable cells were sorted from the digested prostate and further extracted with QIAGEN Rneasy RNA Micro Kit. 150 ng RNA was used for the construction of a sequencing library. For scRNA-seq, the whole prostate was digested by collagenase/hyaluronidase (Cat# 07912) StemCell) for 1-2 h. Viable cells were sorted from whole digested prostate, 10,000 cells were collected for further sequencing library. Sorted viable single cells will loaded into 10xcontroller and individual cells will be captured into oil droplets to form Gel Bead-In-EMulsions (GEMs). The single cell cDNA libraries will be constructed by following 10xGenomics Chromium GEM-X Single Cell 3' v4 Gene Expression User Guide, and then sequenced by illumina Nextseq500 or Novaseq 6000 machine, with targeting minimum 20,000 reads/cell.