title: GSM8737657: sgCdk10_1, replicate 1, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Each sample was placed in a sterile Petri dish on ice. Tumor tissues were transferred into a separate Petri dish and cut into smaller pieces measuring 2–4 mm. The samples were then washed with phosphate-buffered saline (PBS) and further minced into pieces approximately 1 mm³ on ice. Enzymatic digestion was performed using collagenase II (Worthington) at 37°C for 60 minutes with continuous agitation.The resulting cell suspension was passed through a pre-wet 70 µm MACS SmartStrainer filter, and the filtrate was collected into a 50 ml centrifuge tube. The filter was rinsed with 10 ml of RPMI 1640 or DMEM to ensure maximum recovery, and the rinsate was added to the centrifuge tube containing the filtrate. The samples were centrifuged at 300 rcf for 7 minutes at room temperature. After centrifugation, the supernatant was carefully discarded, and erythrocyte lysis was performed immediately.Cell viability was assessed using Trypan blue staining. Finally, an appropriate volume of pre-cooled wash buffer was added, and the cell suspension was gently mixed until the final cell concentration reached 700–1200 cells/µl. Fragmentation, end repair, and A-tailing were performed, followed by purification of the product using SPRIselect beads. Adaptor ligation was then carried out, with subsequent purification using SPRIselect beads. Index PCR was performed, followed by another round of SPRIselect purification. The library concentration was measured using the Qubit庐 3.0 Fluorometer (Life Technologies, CA, USA). The fragment size distribution of the library products was assessed using the Agilent 2100 Bioanalyzer with the High Sensitivity DNA Assay Kit (Agilent Technologies, CA, USA).
identifiers.namespace: GEO
identifiers.id: GSM8737657_r1