title: GSM8650694: VIT010 Patient ATAC; Homo sapiens; ATAC-seq
library_construction_protocol: Human peripheral blood mononuclear cells were isolated by Ficoll-Paque density gradient centrifugation and cryopreserved 2x10e6 cells per sample. Cell viability was 95% or greater for all samples. PBMCs were quickly thawed at 37°C in a water bath and washed with culture medium RPMI 1640 supplemented with 10% FBS. DNase treatment was done before the nuclei isolation. The nuclei isolation was conducted with an RNase inhibitor. Nuclei suspension was filtered with a 40μm cell strainer and counted in a Countess 3 FL Automated Cell Counter (Invitrogen); all samples had <5% live cells. High-quality nuclei were checked by 60x brightfield microscopy. The targeted number of nuclei was 5000 nuclei per sample. Finally, Chromium Next GEM Single Cell Multiome ATAC + Gene Expression kit (10x Genomics) was used to generate single-cell multiome data. CG000338_ChromiumNextGEM_Multiome_ATAC_GEX_User Guide_RevF (Using a microfluidic chip, the nuclei of different patients are partitioned into nanoliterscale Gel Beads-in-emulsion (GEMs) with thousands of Barcodes to separately and uniquely index the transposed DNA and cDNA of each individual nucleus. ATAC and gene expression (GEX) libraries are generated from the same pool of pre-amplified transposed DNA/cDNA and sequenced)