GSM5706742: GEM; Mus musculus; RNA-Seq
After sacrificng mice, tumors were resected from mice, and digested with Dnase, Hyaluronidase, Collagenase. Digested tissues were minced, filtered through a 70-um cell strainer, and made into single-cell suspensions. Single-cell suspensions prepared from Pan02 samples were stained with a Live/Dead fixable dye and sorted by live cells using a BD Aria III sorter. Libraries were prepared using the Chromium controller according to the 10X Single Cell 3' v3 protocol (10x Genomics). The cell suspensions were diluted in nuclease-free water to obtain a cell count of 10,000. The cell suspension was mixed with master mix and loaded with Single-cell 3? Gel Beads and Partitioning Oil into a Single-cell 3' Chip. Transcripts from single cells were uniquely barcoded and reverse-transcribed within droplets. cDNA molecules were pooled, and the cDNA pool was subjected to end-repair, single "A" base addition, and adapter ligation. The products were then purified and enriched via PCR to create the final cDNA library. The purified libraries were quantified using qPCR according to the qPCR Quantification Protocol Guide (KAPA) and qualified using the Agilent Technologies 4200 TapeStation (Agilent Technologies).