GSM9745500: LNP-Sirpa, CD45+ Cells from MC38 Tumors, cDNA, rep2; Mus musculus; RNA-Seq
Tumor tissues were minced into 0.5-mm³ pieces and digested in RPMI-1640 medium with 1% penicillin-streptomycin using the MACS Tumour Dissociation Kit (Miltenyi Biotec) for 30 min at 37 °C with agitation. The digestate was processed as follows: (1) filtration through 70-μm and 40-μm cell strainers (BD Biosciences); (2) centrifugation at 300 × g for 10 min; (3) red blood cell lysis on ice for 2 min; (4) two washes with PBS; and (5) resuspension in PBS with 0.04% BSA. scRNA-seq libraries were prepared with the DNBelab C Series kit (MGI, #940-001818-00). Key steps included: droplet encapsulation, mRNA capture, reverse transcription, cDNA amplification, fragmentation (300-500 bp), and indexed library construction. Library quality was verified with a Qubit ssDNA Assay Kit and Agilent Bioanalyzer 2100. Sequencing was performed on a DNBSEQ-T7 platform (MGI) in paired-end mode: 30 bp (cell barcode/UMI), 100 bp (transcript), and 10 bp (sample index).