GSM7143115: FVB_Dorsal_1; Mus musculus; RNA-Seq
Tissues were minced with razor blades and digested in 0.25% Trypsin-EDTA (Gibco 25200-072) for 10 min at 37°C, followed by incubation for 2.5 hours at 37°C with gentle agitation in DMEM containing 10% FBS, 1 mg/mL Collagenase Type I (Gibco 17100-017), and 0.1 mg/mL of DNase I (Roche 10104159001). Digested tissues were centrifuged at 400 x g for 5 minutes, washed with HBSS, and further incubated in 0.25% Trypsin-EDTA for 10 min at 37°C. Cells were suspended in DMEM containing 10% FBS and 0.4 mg/mL of DNase I and filtered through a 40 μm cell strainer. Libraries for scRNA-seq were prepared using the 10x Genomics Chromium Single Cell 3' Library and Gel bead Kit V2 (CG00052_RevF) according to the manufacturer's protocol for each dissected mouse prostate lobe and each human prostatectomy tissue punch. The cDNA libraries were sequenced (150 bp paired-end) on the Illumina HiSeqX platform. Each sequenced library was demultiplexed to FASTQ files using Cell Ranger (10x Genomics).