Name: GSM6052782
Instrument: Illumina NovaSeq 6000
Strategy: ATAC-seq
Source: GENOMIC
Selection: other
Layout: PAIRED
Construction protocol: The mouse fetus at desired dates was isolated using cesarean operation when the pregnant mice were anesthetic with the compound ketamine. The skin tissues (0.5 cm × 0.5 cm) were isolated from the fetus back skin and were immediately transferred to the ice-cold DMEM/F12 media (Gibco, Beijing, China) with 50 U/ml penicillin and 50 mg/ml streptomycin (HyClone, Beijing, China). After washing three times with DMEM/F12 to remove contaminative blood cells, the skin tissues were then dissociated into single cells prior to sequencing. For E13.5 and E16.5 skin tissues, the obtained skin tissues were firstly incubated with 2 mg/ml collagenase IV (Sigma, St Louis, MO, USA) for 30 min at 37 °C, and then the skin tissues were mechanically dissociated into single-cell suspensions with a 1ml pipette tip. For P0 skin tissues, the skin tissues were firstly cut into ~3 mm skin pieces and then incubated with 2 mg/ml collagenase IV for 30 min. After incubation, the hair follicles within the skin tissues were isolated with a pair of precise forceps and the pooled hair follicles were further dissociated into single cells with TypLE Express (Gibco, Grand Island, NY, USA) for 30 min at 37 °C. The obtained single-cell suspensions were then washed three times with PBS supplemented with 0.04% BSA (Sigma, St Louis, MO, USA) and were filtered with a 40 μm cell strainer (BD Falcon, BD Biosciences, San Jose, CA, USA) to remove debris and cell aggregations. For each stage, the samples were obtained from at least 3 different mouse fetus and for each fetus mouse, the single-cell suspension was prepared separately until finally pooled together prior to single-cell barcoding. Single-cell ATAC-seq libraries were prepared using 10x Chromium Single Cell ATAC Library & Gel Bead Kit (10x Genomics Inc., Pleasanton, CA, USA) following the manufacture's introduction.