title: GSM7510838: 506_Pb-PRL_ATT; Mus musculus; RNA-Seq
library_construction_protocol: The prostates were dissected and minced, then digested for one and a half hours with 1mg collagenase at 37°C in 10 ml DMEM + 10% SVF. The cells were then dissociated with trypsin and sieved using a 40µM cell sieve, then cleared of debris using histopaque centrifugation. Live ter119-negative cells were then isolated by flow cytometry. The scRNA-seq libraries were generated using Chromium Single Cell Next GEM 3′ Library & Gel Bead Kit v.3.1 (10x Genomics) according to the manufacturer's protocol. Briefly, cells were counted, diluted at 1,000 cells/µL in PBS+0,04% and 20,000 viable cells were loaded in the 10x Chromium Controller to generate single-cell gel-beads in emulsion. After reverse transcription, gel-beads in emulsion were disrupted. Barcoded complementary DNA was isolated and amplified by PCR. Following fragmentation, end repair and A-tailing, sample indexes were added during index PCR. The purified libraries were sequenced on a Novaseq (Illumina) with 28 cycles of read 1, 8 cycles of i7 index and 91 cycles of read 2.