GSM7730907: gom,bamapig,snRNA_seq; Sus scrofa; RNA-Seq
The quality control standards for preparing single-cell suspension samples encompassed specific criteria: a cell viability threshold surpassing 80%, a cell concentration range of 700 to 1200 cells/μL, and cell diameters within 5 to 30 μm. For the PMM, SAT, and GOM samples, cell nuclei extraction took place during sample preparation using the 10x Chromium Nuclei Isolation Kit due to their notably enlarged cell diameters. Ensuring the viability of cell/cell nucleus suspensions, which met the set standards, these were subsequently loaded onto 10x Genomics Single Cell 3.0 Chips.During this loading step, cells were partitioned into Gel Beads-in-Emulsion (GEMs) along with gel beads coated with 10x barcode oligonucleotides, encompassing a 14-bp index and a 10-bp UMI. Following GEMs generation, the samples were transferred to PCR tubes for reverse transcription, a step executed employing a T100 Thermal Cycler (Bio-Rad).