title: IE11.5 male
library_construction_protocol: Single-cell suspensions were prepared from testes from three E14.5 Nanos2-KO embryos by incubation with 0.15% trypsin-EDTA at 37C for 5-10 min. After incubation, wash medium was added to inhibit trypsin and cells were filtered through a 35-um sieve (BD Bioscience). Cells were re-suspended in DMEM containing 10% FBS and the cell density was adjusted to 1.0x10^6 cells/ml. We tried to load 3,000 cells on the Chromium Controller (10x Genomics Inc.) and constructed the scRNA-Seq libraries using Chromium Single Cell 3' Reagent Kits v2 following the manufacturer's instructions. Each library was read at a depth of 1 million per sample for cell number estimation and 100 million per sample for analysis using Hiseq2500.