GSM7660360: Hunner_Type_Interstitial_Cystitis_Case8_Hunner_Lesion; Homo sapiens; RNA-Seq
Total RNA was extracted from frozen samples using NucleoSpin RNA (Macherey-Nagel, Düren, Germany). RNA was quantified on a NanoDrop 2000c Spectrophotometer (Thermo Fisher Scientific, San Jose, CA, USA), and the purity was measured using an RNA ScreenTape assay on a 2200 TapeStation (Agilent Technologies, Santa Clara, CA, USA). Quality assessment was performed using 2200 TapeStation Software (Agilent Technologies). Purified total RNA (10 ng) was resuspended in Clontech buffers for mRNA amplification using 5′ template switching PCR with a Clontech SMART-Seq v4 Ultra Low Input RNA kit (Clontech, Palo Alto, CA, USA) according to the manufacturer's instructions. Amplified cDNA was fragmented and appended with dual-indexed barcodes using Illumina Nextera XT DNA Library Prep kits (Illumina, San Diego, CA, USA).