GSM8527855: Human_Rep1; Homo sapiens; RNA-Seq
Renal biopsies were minced into small pieces and were digested in 1 mL HBSS (Ca/Mg) including DNAse I (10mg/mL) and Collagenase I (100 U/µL) at 37°C and 350 rpm for 30 min. Addition of 2ml PBS to all organs was followed by filtering suspensions trough a 70 µm cell strainer. Samples were then centrifuged at 400 xg for 5 min and cell pellets were resuspended in PBS with 1% FCS. Approximately 2,000-3,500 vital cells of each sample were processed for scRNA-seq using using BD Rhapsody Targeted mRNA & AbSeq Amplification Kit (BD Biosciences, Cat. No. 633774) according to the manufacturer's instructions. Quality, purity, average size and concentration of copy DNAs and libraries were determined by measurements on a Qubit FLuorometer (Thermo Fisher Scientific) and a Tapestation 2200 (Agilent Technologies). Indexed libraries were sequenced using a NovaSeq 6000 sequencing platform with PE150. flow cell and reagents (Illumina).