GSM7900393: Single cell multiome 10x from lineage-Sca1+ckit+ sorted marrow T58A cells replicate 1 ATAC; Mus musculus; OTHER
Single celll multiome analysis ( scRNA-Seq and scATAC-Seq) was done using nuclei isolated according to the manufacturers specifications (10X Genomics). For this, cells were sorted on a flow cytometer (Lin-Sca1+ckit+), and nuclei were prepared in buffer containing digitonin. Cells were counted, barcoded, and extraction was carried out in the Fred Hutch Cancer Center genomics core as described for the 10X genomics system. After barcoding and extraction, single cell multiome libraries were constructed in the Fred Hutch Cancer Center genomics core using the 10X Genomics system. RNA-seq libraries were prepared from total RNA using the TruSeq RNA Sample Prep v2 kit. For CUT&RUN, cells were processed on a previously published, automated system at the Fred Hutch Cancer Center that uses KAPA DNA polymerase method for library preparation optimized for recovery of small DNA fragments.