title: GSM8793322: duodenum_WT; Mus musculus; RNA-Seq
library_name: GSM8793322
library_construction_protocol: 2 cm of duodenum was collected from mice with the indicated genotype. After washing with ice-cold PBS, the tissue was minced into 2-3 mm pieces, washed twice with ice-cold PBS, digested with digestion buffer [2 mg/mL Collagenase II (Sigma), 0.05 mg/mL Dnase I (Solarbio), and 2 mg/mL Dispase (Worthington)] at 37℃ with 100 rpm for 40 min, and stopped with three times the volume of ice-cold RPMI 1640 (Gibco). Cell suspensions were passed through a 70 µm filter, and centrifuged at 350 g for 5 min at 4℃. The supernatant was carefully removed, the cell pellets were collected and red blood cells were removed using 1 ml Red Blood Cell Lysis Solution. Then they were resuspended in 1× DPBS (0.04% BSA) and centrifuged twice at 300 g for 10 min at 4℃. Single-cell suspensions were counted using a cell counter (Countstar), and the final concentration was adjusted to 700-1200 cells/μL. The single-cell suspensions were loaded on a 10X Genomics GemCode Single-cell instrument to generate single-cell Gel Beads-In-EMulsion (GEMs). Libraries were generated and sequenced from the cDNAs with Chromium Next GEM Single Cell 3′ Reagent Kits v3.1.