GSM5242819: Zfp541-KO whole testis; Mus musculus; RNA-Seq
Droplet-based scRNA-seq (10x Chromium) was performed by Genergy Bio-Technology (Shanghai) Co., Ltd. Testicular tissues were disassociated enzymatically using enzyme mixtures comprising of 1 mg/mL collagenase IV (GIBCO,17104-019), 1.5 mg/mL hyaluronidase (SIGMA，H2126), 1 mg/mL trypsin (SIGMA，T1426), and 1 mg/mL DNase I (SIGMA, DN25), dissolved in DMEM medium at 35℃ with gentle agitation for 20 mins. Subsequently, the cell suspension was filtered through a 40 μm cell strainer, centrifuged at 250×g for 5 min at 4 °C, the supernatant was removed and cells in the pellet were resuspended in cold PBS (Phosphate buffer saline) containing 0.05% BSA carefully. Tetraploid germ cell populations were then isolated by flow cytometry as previously described (Jung et al., 2019). Single tetraploid germ cells were stained with trypan blue (Invitrogen) and counted using light microscopy and then loaded onto a 10x Chromium microfluidics system according to the manufacturer's guidelines. Libraries were generated using Single Cell 3′ library and Gel Bead Kit v.3 according to the protocol guidelines. The final library pools were sequenced on an Illumina Novo 6000 platform instrument using 150-base-pair paired-end reads.
GEO Accession
GSM5242819