GSM8993331: Normal_2; Oryctolagus cuniculus; RNA-Seq
A 2.0×2.0 cm area of mucosa was extracted from rabbit bladder, then finely cut into 1mm^3 pieces with blade and digested with dissociation buffer (Ham's F12, 1 mg/ml collagenase Ⅱ (Gibco), 100 µg/ml Pen/Strep) for 1-2 hour at 37 °C with gentle rocking. Dissociated cells were washed thoroughly in cold wash buffer (Ham's F12, 5% fetal bovine serum, 100 µg/ml Pen/Strep) before proceeding to scRNA preparation. The single cell suspensions from normal and injured bladder tissues were procceded to scRNA-seq, according to instruction manual of the Chromium Single Cell 3' Reagent Kits v3. Cellular suspensions were loaded on the Chromiu Controller (10x Genomics, Pleasanton) to generate Gel Bead-In-Emulsions (GEMs). Barcoded sequencing libraries were conducted following the instruction manual of the Chromium Single Cell 3' Reagent Kits v3 (10x Genomics). Following the library preparation, the sequencing was performed with paired-end sequencing of 150nt each end on one lane of NovaSeq 6000 per sample.