GSM9461542: E18.5_nasal_sample2; Mus musculus; RNA-Seq
The tissues were digested in 10 U/mL Papain (Worthington, LS003126) for 10 min at 37 °C, 5% CO2 incubator, and then cut approximately 50 times. 100 U/mL DNase I (Worthington Biochemicals, LS006343) was added and the mixture was incubated for an additional 5 to 10 minutes. The enzymatic reaction was then terminated, followed by supplementation with 20 U/mL DNase I to ensure complete genomic DNA digestion. After digestion, cells were filtered through 100-µm cell strainers (Falcon, 352360) on ice and centrifuged at 4 °C, 300× g for 5 min. Single-cell suspensions were loaded onto 10× Genomics Chromium system to generate single-cell gel beads-in-emulsion (GEMs). After incubation, GEMs were disrupted and cDNA was amplified via PCR. For library preparation, 25% of total cDNA (10 μL) was used to construct 3' gene expressioen libraries, which were then purified with SPRIselect beads.