GSM9080119: Aged kidney treated with BPTES, sample 1; Mus musculus; RNA-Seq
Kidneys were minced by a sharp blade and digested with liberase solution (50 μg/ml liberase TH, 2 Kunitz units/ml DNase I, and 10 mM HEPES in RPMI) at 37oC for 1 hr. During the digestion, the tissue suspensions were passed through 18 G and 21 G needles sequentially to further homogenize residual tissues. After the digestion, all samples were passed through 70-μm cell strainer and pelleted by centrifugation at 400g for 5 mins. After RBC lysis and FcR blocking, cells were stained with anti-CD45 on ice for 30 mins. 20,000 CD45- and 9,000 CD45+ cells from the same tissue were mixed for preparing one cDNA sample. The library with dual index was performed according to the manufacturer's instructions (Chromium GEM-X Single Cell 3' Reagent Kits v4, 10x Genomics).