title: GSM9464503: CON_2; Rattus norvegicus; RNA-Seq
library_construction_protocol: For snRNA-seq, ileal tissue (~100 mg) was processed for nuclei isolation using the Boyou® Cell Nucleus Isolation Kit (SHBIO, 52009-10, China) according to the manufacturer's protocol. Nuclei were counted and quality assessed using a fluorescence cell analyzer (Countstar® Rigel S2/Mira FL) and microscopy. For spatial transcriptomics, ileal tissues were embedded in OCT within 30 min after resection, snap-frozen on dry ice, stored at −80°C, sectioned at 10 μm on a cryostat, and mounted onto Stereo-seq chips for downstream permeabilization and reverse transcription. snRNA-seq libraries were prepared using the DNBelab C High-throughput Single-cell RNA Library Preparation Kit V3.0 (MGI Tech). Single nuclei were co-encapsulated with barcoded gel beads and reverse transcriptase mix into droplets using the DNBelab C-TaiM 4 system, followed by cDNA synthesis, emulsion breakage, cDNA purification and amplification, fragmentation, end repair/A-tailing, adaptor ligation, size selection (300–600 bp), PCR amplification, and library purification.