GSM4844219: H3K27me3_Rep1; Mus musculus; ChIP-Seq
Basal layer epidermal progenitors were isolated from K14-RFP;K10-H2B-GFP mice. The basal layer epidermal progenitors were isolated from adult back skin. Back skins were collected and washed with 1x DPBS and incubated for 0.25% Trypsin/EDTA at 37℃ for 1 hour. Crosslinking was stopped by the addition of Glycine (final concentration 125mM) for 5 minutes at room temperature, followed by 2 washes with 1x DPBS.  Cells were stained with 1:200 Sca1-PerCP-Cy5.5 (Biolegend) and Zombie violet (Biolegend) for 30 minutes on ice and washed twice with 1x DPBS before cell sorting. Epidermal progenitors were sorted as Sca1(+), RFP(high) and GFP(-). For high-throughput ChIP sequencing, libraries were constructed from 5ng of purified DNA using the NEBnext Ultra II DNA Library Prep Kit (New England Biolabs) according to the manufacturer's instructions. Constructed ChIP-seq libraries were sequenced on Illumina NextSeq 500 platform with 75-single end reads.