title: GSM7720655: day 2; Homo sapiens; RNA-Seq
library_construction_protocol: All protocols are same as in GSE175525.  The suspension of cells were generated as the same manner with passage or the induction of differentiation. For the aggregates of PGCLC induction at day 0.5, 1, 1.5 and 2, the aggregates were collected, washed once in PBS, and dissociated with 0.25% Trypsin–EDTA for 10–15 min at 37°C with gentle pipetting every 5 min. Trypsin was neutralized with a 5× volume of 10% FBS in DMEM and washed once with 0.1% BSA in PBS.  Prior to proceeding the scRNA-seq analysis, the live single cells were sorted using FACS Aria III system (BD Biosciences) by FSC-SSC and DRAQ7 (ab109202) staining for iMeLC, or FSC-SSC and EGFP-tdTomato gating with DRAQ7 staining for day 0.5, 1, 1.5 and 2 PGCLCs. Single-cell RNA-seq libraries of 10X data were generated using the 10X Genomics Chromium Controller (10X Genomics) and Chromium Single Cell 39 Reagent Kits v3.1 according to the manufacturer's instructions.  The final libraries were quantified using a KAPA library quantification kit (KK4824), and the fragment size distribution of the libraries was determined using a LabChip GX DNA high sensitivity kit (Perkin Elmer).  Pooled libraries were then sequenced using a NovaSeq 6000 Illumina Sequencer with a NovaSeq 6000 S1 Reagent Kit (100 Cycles, 20012865).