title: GSM9209748: UC-P12 Organoid; Canis lupus familiaris; RNA-Seq
library_construction_protocol: For single-nuclei samples, frozen tissues were received at Azenta, South Plainfield, NJ, USA in dry-ice and stored in liquid nitrogen until further processing. Nuclei extraction was performed using the Miltenyi Nuclei Extraction Buffer (Miltenyi Biotec, Auburn, CA, USA) following manufacturer's guidelines with gentle MACS Dissociation and C tubes. Upon isolation, the nuclei were counted using AO/PI dye on the Nexcelom Cellaca MX before loading onto the Chromium Controller. Single-nuclei RNA libraries were generated using the Chromium Single Cell 3' kit (10X Genomics, CA, USA). Loading onto the Chromium Controller was performed to target capture of ~6,000 GEMs per sample for downstream analysis and processed through the Chromium Controller following the standard manufacturer's specifications. The sequencing libraries were evaluated for quality on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA). Libraries were quantified using qPCR (Applied Biosystems, Carlsbad, CA, USA) prior to loading onto an Illumina NovaSeq XPlus instrument. The samples were sequenced at a configuration compatible with the recommended guidelines as outlined by 10X Genomics. Raw sequence data (.bcl files) were converted into fastq files and de-multiplexed using the 10X Genomics' cellranger mkfastq command.