GSM8214794: DT D2; Mus musculus; RNA-Seq
Fresh gastric corpus was harvested and mechanically dissociated in DMEM media+ Collagenase 1A while shaking at  37° for 30 minutes. Tissue was further dissociated chemically in 5-15 minutes at 37° in 0.05% trypsin-EDTA. The single cell suspension was passed through a 30µm filter, quantity and viability assessed with hematocytometer, and submitted to the Single Cell Genomics Core at Baylor College of Medicine in 0.04% BSA 1x PBS. The single-cell gene expression Library was prepared according to the Chromium Single Cell Gene Expression 3'v3.1 instruction (PN-1000121, PN-1000120, PN-1000213, 10x Genomics). Briefly, single cells, reverse transcription (RT) reagents, Gel Beads containing barcoded oligonucleotides, and oil were loaded on a Chromium controller (10x Genomics) to generate single-cell GEMS (Gel Beads-In-Emulsions) where full-length cDNA was synthesized and barcoded for each single cell. Subsequently, the GEMS are broken and cDNA from each single cell is pooled. Following cleanup using Dynabeads MyOne Silane Beads, cDNA is amplified by PCR.  The amplified product is fragmented to optimal size before end-repair, A-tailing, and adaptor ligation. The final library was generated by amplification. After passed the quality control, the next-generation sequencing of libraries was performed on NovaSeq 6000 (Illumina).