GSM4973574: HemaCare-D273097 (10X); Homo sapiens; ATAC-seq
Nuclei were isolated from fresh pancreatic islets using a glass dounce homogenizer with a tight-fitting pestle, frozen whole pancreas using a mortar and pestle, and PBMCs using a lysis buffer. (CB; combinatorial barcoding) Permeabilized nuclei were distributed to 96 wells and tagmentation was carried out to introduce a first barcode combination. After pooling, 20 nuclei were sorted into each well of a 96 well plate and a second barcode was introduced by PCR. Libraries were purified according to the MinElute PCR Purification Kit manual using a vacuum manifold and size selection was performed with SPRI Beads. (10X; 10X Genomics) Libraries were generated using the 10X Genomics Chromium Chip E Single Cell ATAC kit following the manufacturers instructions.