GSM8255580: GF_NCD, replicate1, scRNA-seq; Mus musculus; RNA-Seq
Peyer's patches dissected from mice were minced with scissors and then stirred in a 20-mL conical flask containing 10 mL of RPMI medium supplemented with 2% (v/v) FCS, 1 mg/mL Collagenase (WAKO), and 50 μg/mL DNase, using a stir bar at 37°C and 500 rpm for 15 minutes. The suspension was then passed through an 18 G needle 10 times and stirred at 37°C and 500 rpm for an additional 15 minutes. Samples were filtered through a 100-μm Cell Strainer, centrifuged at 4°C and 500 g for 5 minutes, and CD19-negative PI-negative cells were sorted by FACS for subsequent scRNA-seq analysis. Library was prepared using Chromium Single Cell 3' Library & Gel Bead Kit v2 (10x Genomics) according to the manufacter's instructions.