GSM8606253: 94BLA, Unpaired-encoding, rep 2, ATAC; Rattus norvegicus; ATAC-seq
single nuclei from the tissues were isolated using the 10x Genomics Chromium Nuclei Isolation with RNAse Inhibitor Kit (PN-1000494). The tissues were immediately immersed in pre-chilled lysis buffer post-extraction to facilitate dissociation and nuclei release. Nuclei were isolated by passing the suspension through a 40 μm cell strainer (Bel-Art™) to remove clumps and aggregates. The quality and concentration of nuclei were assessed by staining with a Trypan blue and ethidium bromide mixture, using a hemocytometer (NanoEntek) and fluorescence microscopy (Invitrogen™ EVOS™ M5000). The final nuclei concentration was optimized by diluting the nuclei buffer. Single-nucleus ATAC and RNA-seq libraries were constructed using the 10X Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Kit (10x Genomics, User Guide Rev F, CG000338). Approximately 2,000 nuclei per sample were loaded onto the Chromium Next GEM Chip G. Libraries were prepared in parallel for ATAC-seq and RNA-seq, comprising double- strand DNA with standard Illumina paired-end construction. Sequencing was performed on an Illumina NovaSeq 6000 platform (Sickkids, Canada) to achieve a target depth of ~20,000 read pairs per cell for ATAC-seq and ~25,000 read pairs per cell for RNA-seq.