title: GSM8316751: FGC_E14.5_0513; Mus musculus; OTHER
library_construction_protocol: Eight- to ten-week-old wild-type female DBA/2N mice in natural estrus were mated with male C57BL/6J (Oct4-EGFP) transgenic mice. When female mice with evident vaginal plug around noon, the day was set as E0.5. Oct4 positive FGCs were isolated from E11.5, E13.5, E14.5, E15.5, E16.5, E17.5 and E18.5 fetal gonads by FACS. At E13.5-E18.5, male and female gonads could be distinguished from morphology which based on whether the testicular cords were presence or not, and gonads cells were collected with male or female respectively. Whole gonads at different stages were digested using the 0.25% trypsin-EDTA for 30 min at 37 °C, and then added 0.25% BSA to stop digestion. Gonads cells were centrifuged at 2,000×g. A total amount of 0.5μg to 2 μg  DNA per library was used as input material for the DNA library preparations. SQK-LSK109 Kit (Nanopore, UK) was used to construct 1D library. The DNA library was constructed by standard ligation method without DNA fragmentation, after end repaired, added the sequencing adaptor, motor protein and tether protein were connected to prepare the DNA library. After that, each amplicon fragment library was loaded into one R9.4.1 or R10.4.1 flow cell and sequenced on PromethION 48 model.