GSM4476449: SPGI_wtLM_S1_2to4C_DMRT1hi_STRA8.mm10.RT; Mus musculus; OTHER
Nuclei from mouse or human testes were prepared as described in (Lam et al., 2019). To purify the populations of interest, a combination of intranuclear markers were used (Table 1, Figure 2). Pure nuclei populations were resuspended in 300 ul of lysis buffer (SDS 1%, 10mM EDTA, 50mM, Tris pH=8) and then sonicated in a Bioruptor UCD200 for 20 min 30 sec ON 30 sec OFF, “high” setting. NaCl was added to a final concentration of 0.2M and the sample incubated overnight at 65C to reverse DNA-protein crosslinks. 5U of Proteinase K (NEB Cat. No. P8107S) was added and the sample further incubated for 1h at 45C. DNA was purified with MinElute columns (QIAGEN), as indicated by the instruction manual. Whole genome sequencing libraries were prepared from 100 ng of purified DNA using the KAPA Hyper Prep Kit (Cat. No. KK8502) following the manufacturer's instructions.