GSM7502562: Lpar6_KO, Lpar6 knockout, scRNAseq; Mus musculus; RNA-Seq
The bladders were harvested after ligating the urethra and injecting 50 µl of 0.25% Trypsin in PBS. The cells were incubated in Trypsin at 37°C for 30 minutes. The urothelial cell clusters and tissue fragments were removed by passing the cell suspension through the 70µm cell strainer. The red blood cells were removed by Ficoll gradient centrifugation. The collected single cell suspensions were washed twice with PBS containing 0.04% BSA and resuspended in 200µl of PBS. Library preparation steps were carried out by running ThermoFisher Qubit HS dsDNA Assay along with Agilent HS DNA Bioanalyzer for concentration and quality assessments. Equal amounts of each uniquely-indexed sample library were pooled together. The resultant pool was verified for concentration via qPCR using a KAPA Biosystems KAPA Library Quantification Kit.