GSM8338491: Group B, replicate 4, snRNAseq; Homo sapiens; RNA-Seq
After being removed from the -80°C refrigerator, the heart samples were dissected in 4°C PBS buffer (#C10010500BT, Gibco, USA). The tissue fragments were resuspended in 30ml lysis buffer. The suspension fluid was homogenized in the Dounce tissue grinder set (Pestle B). The supernatant was filtered through 100μm and 70μm filters. After centrifugation (700rcf, 10 min, 4°C), the supernatant was discarded and the pellet was resuspended using 20 ml preservation buffer. After recentrifugation (13000rpm, 1h, 4°C), the supernatant was discarded, and the pellet was resuspended using PBS buffer. The nuclei suspension was mixed with trypan blue (1:1) and counted by using the hemocytometer. The processing of libraries was performed according to the manufacturer's instructions. The snRNA-seq libraries were prepared using the ChromiumTM Next GEM Single Cell 3' GEM, Library & Gel Bead Kit, version 3.1, 16 reactions (10X Genomics) and sequenced using the NoveSeq 6000 (Illumina, USA).