E13.5 testes were digested in 0.05% Trypsin-EDTA and immunostained with antibodies to PE-PECAM1 (Biolegend, 102407)and APC-c-Kit (Biolegend, 313205). Germ cells were identified on the basis of OCT4-GFP+ and further separated into AP (PECAM1-high;cKit-high) or MD (PECAM1-low;cKit-low) populations. Cells were lysed using EZ DNA Methylation-DirectTM Kit (Zymo D5020). Bisulfite conversion and library preparation was carried out with the Pico Methyl-Seq Library prep kit (Zymo D5455). Libraries were sequenced on NextSeq 500 High Output and NovaSeq 6000 S1 flow cells 484 yielding to 75-bp, and 50-bp paired-end reads, respectively for whole-genome bisulfite sequencing by post-bisulfite adapter tagging.
GEO Accession
GSM4695833