title: GSM8636837: Control; Mus musculus; RNA-Seq
library_name: GSM8636837
library_construction_protocol: Tumor tissues were harvested from the tumor-bearing mice. Tumor tissues were digested using a tumor dissociation kit and dissociated into single cells using a gentleMACSTM Octo Dissociator. Red blood cells (RBCs) in a single-cell suspension derived from tumor tissues were lysed using RBC lysis buffer, and debris and dead cells were removed using a debris removal solution and dead cell removal kit. Viable cells were counted and processed to generate an RNA library following the 10X Chromium Single Cell V(D)J User Guide (CG000086). Briefly, cell suspensions were diluted in nuclease-free water to achieve a target cell count of 10,000 cells. The suspension was combined with a master mix and loaded onto a Single-Cell-A-Chip together with Single-Cell 5′ Gel Beads and Partitioning Oil. Within individual droplets, the RNA transcripts from single cells were uniquely barcoded and reverse-transcribed. The resulting cDNA molecules were pooled and PCR-enriched. For the V(D)J Enriched Library, the cDNA pool was first amplified using the T Cell Mix1 or B Cell Mix1 primer, followed by a second amplification using the T Cell Mix2 or B Cell Mix2 primer. For the 5′ Gene Expression Library, the cDNA pool underwent an end repair process, the addition of a single A base, and subsequent adapter ligation. The resulting products were purified and PCR-enriched to create the 5′ Gene Expression Library. The purified libraries were quantified using qPCR according to the KAPA qPCR Quantification Protocol Guide, and the quality was checked using an Agilent Technologies 4200 TapeStation. Sequencing was performed on a HiSeq X Ten (Illumina) following the recommended read-length guidelines.
identifiers.namespace: GEO
identifiers.id: GSM8636837_r1