title: GSM8241034: xrTestis_d127_human; Homo sapiens; RNA-Seq
library_construction_protocol: (SINGLE CELL) xrTestis-derived cells were sorted via FACS for expression of AG/VT/PC and collected in CELLOTION. Cell clumps were removed by straining cell suspensions through a 70 µm nylon mesh and then the single-cell suspension was centrifuged at 300 × rcf for 5 min. Cells were resuspended in 0.1% BSA in PBS and counted. All samples had greater than 80% viability as determined by trypan blue staining. Encapsulation was performed using a 10X Genomics Chromium Controller according to the manufacturer's protocol. (WGBS) PGCLC aggregates were digested with 400 μl 0.25% trypsin for 15 min. Then, 100 μl FBS was used to stop digestion, and the lysates were pipetted well to obtain a single-cell suspension. The fractions of TFAP2C-eGFP, TFAP2C-eGFP+DDX4-tdTomato and DDX4-tdTomato were sorted for methylome analyses. Cells were collected and lysed in 50 mM Tris (pH 8.0), 10 mM EDTA, 0.5% SDS, and 100 μg/ml proteinase K. (SINGLE CELL) The Cells were loaded into chromium microfluidic chips with the Chromium Single Cell 3ʼ Reagent Kit (v3.1 chemistry) and then used to generate single-cell gel bead emulsions (GEMs) using the Chromium Controller (10× Genomics) according to the manufacturer's protocol. reverse transcription step was performed using a C1000 Touch Thermal Cycler with 96-Deep Well Reaction Module (Bio-Rad). All subsequent cDNA amplification and library construction steps were performed according to the manufacturer's protocol. (SINGLE CELL) Chromium Single Cell 3ʼ Reagent (v3.1 chemistry) was used for library preparation (WGBS) Crude DNA was used to build a sequencing library according to the protocol of the Pico Methyl-Seq Library Prep Kit (Zymo, #D5455). The libraries were sequenced on the Illumina 2200 platform.