title: GSM8471374: Sca1+ FAP_Kdm6b-CutNtag_WT_rep1; Mus musculus; OTHER
library_name: GSM8471374
library_construction_protocol: Single cells were collected from mouse muscle by disection and mincing of muscle tissue, digestion using collagenase II and dispase in wash medium, and training using a 20-gauge needle and a 40 micrometer filter. FAPs were then isolated by Sca1+, propidium iodide negative FACS sorted cells. About 10,000 of FAPs (Sca1+) from WT, Mdx, and Utrn-/Mdx mice were collected by FACS. FAPs were bound with concanavalin A beads for immobilization, then incubated with primary antibodies (Kdm6b, H3K27me3, Fosl1) at 4 ℃ overnight and secondary antibody 30 mins at RT. After, cells are immobilized with concanavalin beads, permebilized cells were tagmented with pA-Tn5 with sequencing adapter. This is then followed by DNA purification, and final libraries were amplified by Q5 polymerase (NEB, M0491L) and quantified using a Qubit™ 1X dsDNA HS Assay Kit and a High Sensitivity D1000 Tapestation.
identifiers.namespace: GEO
identifiers.id: GSM8471374_r1