GSM9213642: Sample8_Gut3KL; Mus musculus; RNA-Seq
To obtain single cell suspensions, peeled ME was cut in 1-2 mm pieces and digested with 2 mg/mL Collagenase type IV and 0.8 mg/mL dispase in RPMI (Gibco) supplemented with 2% HEPES (Gibco), 2% FBS (Gibco), and 50μg/mL DNase for 1h at 37°C with continuous agitation. The single cell suspensions were then homogenised with a potter grinder and strained through 70 μm cell strainers (BD Falcon). Single-cell libraries were prepared using 10X Genomics Chromium Next GEM Single Cell 5′ kit v3.1, according to the manufacturer's instructions. Libraries were sequenced using an S2 flowcell on an Illumina NovaSeq S6000 instrument, with sequencing parameters recommended by 10X Genomics, aiming for 40,000 reads per cell.