GSM7179777: hematopoietic stem and progenitor cells, young mice fed ad libitum , scATACseq; Mus musculus; ATAC-seq
We were able to isolated 50,000 hematopoietic stem and multipotent progenitor cells from each mouse using antibody staining and FACS. Cell suspensions from two mice were pooled for nuclei isolation. Nuclei Isolation for Single Cell Multiome ATAC + Gene Expression Sequencing was performed according to protocol (PROTOCOL VERSION, lysis time 3 minutes). Nuclei suspensions were loaded into separate channels of the Chromium Controller (10x Genomics) and processed with Single Cell Multiome ATAC + Gene Expression (v1 chemistry) following the manufacturer's protocol (Document Number CG000338 Rev E).     ATAC libraries: 7 cycles in sample index PCR reaction and evaluated using D5000 ScreenTape (Agilent 4200 TapeStation System) and DNA 7500 (Agilent 2100 Bioanalyzer).   Gene Expression libraries: cDNA was amplified in 7 cycles, total cDNA was assessed on high sensitivity DNA assay (Agilent 2100 Bioanalyzer), another 6 cycles of sample index PCR were performed and final libraries were evaluated using D5000 ScreenTape (Agilent 4200 TapeStation System).     Each type of library was pooled and sequenced using Illumina NovaSeq 6000 platform and two SP Reagent Kits v1.5 (100 cycles). scATAC-seq: Read 1 and Read 2 53 bp for DNA Insert, i7 8 bp for sample index and i5 24 bp for 10x Barcode and Spacer. scRNA-seq: Read 1 28 bp for 10x Barcode and UMI; Read 2 90 bp for Insert, i7 and i5 10 bp for sample index.