GSM8199187: te2_2; Mus musculus; RNA-Seq
Paired ventral prostate lobes were collected into one tube for each animal in ice-cold PBS and were transferred to 100 ul of 10 mg/mL (in HBSS) Bacillus licheniformis protease (Creative Enzymes, Shirley, NY). Tissues were agitated at 6°C at speeds ranging from 600 to 1000 rpm for up to one hour, with intermittent pipetting, until noticeable cell separation was observed. Cells were then passed through a 40µm mesh strainer and cell viability was assessed on a Countess 3 FL and ranged between 72-78 %. Following the guidelines from the Chromium Next GEM Single Cell manual, we determined the optimal cell loading to be 7000 cells per sample. Cells were barcoded and libraries were generated for duplicate samples per group using the Chromium Next GEM Single Cell 3ʹ Kit v3.1. Libraries were created from each animal and concentrations were calculated based on Tapestation data (200-1000 bp). Paired-end sequencing was conducted on a NextSeq2000 instrument.