GSM8844020: AR4N (3'); Homo sapiens; RNA-Seq
Samples were washed, chopped into small pieces, and digested in an enzymatic digestion buffer containing 3 mg/ml collagenase type II (#LS004176, Worthington Biochemical Corp, NJ, USA), 0.15 mg/ml collagenase type XI (#H3506, Sigma Aldrich, MO, USA), 0.25 mg/ml soybean trypsin inhibitor (#LS003571, Worthington Biochemical Corp), 0.1875 mg/ml elastase lyophilized (#LS002292, Worthington Biochemical Corp), 0.24 mg/ml hyaluronidase type I (#H3506, Sigma Aldrich), and 2.38 mg/ml 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid (HEPES, #H4034, Sigma Aldrich) in PBS with Ca2+ and Mg2+, at 37°C for 90 minutes with shaking. The digested samples were filtered through 40 μm cell strainers (#352340, BD Biosciences, NJ, USA), washed, and centrifuged at 3000 × g for 10 minutes. ScRNA-seq (3') libraries were prepared using the Single Cell 3'Reagent Kits v3 for scRNA-seq (10x Genomics, CA, USA). Sequencing libraries were generated following the User Guide (CG000388).