GSM2773166: 234_kidney_1_ccpr; Homo sapiens; RNA-Seq
Total RNA was extracted using QIAzol lysis Reagent and miRNAeasy Mini Kits (QIAGEN, Hilden, Germany), following Qiagen protocol. Addition of wash buffers RPE and RWT (QIAGEN) was done mechanically by using the QIAcube (QIAGEN, Hilden, Germany) via the miRNAeasy program. Isolated RNA was and stored at −80 °C ERCC RNA Spike-In Control mix 1 (Thermo Fisher Scientific) was added to the total RNA and Poly(A) RNA was prepared using the Dynabeads mRNA DIRECT Purification Kit (Thermo Fisher Scientific). Bar-coded RNA libraries were generated according to the manufacturers’ protocols using the Ion Total RNA-Seq Kit v2 and the Ion Xpress RNA-Seq bar coding kit (Thermo Fisher Scientific). The size distribution and yield of the barcoded libraries were assessed using the 2200 TapeStation System with Agilent D1K ScreenTapes (Agilent Technologies). Sequencing templates were prepared on the Ion Chef System using the Ion PI IC 200 Kit (Thermo Fisher Scientific). Sequencing was performed on an Ion Proton System using Ion PI v2 chips (Thermo Fisher Scientific) according to the instructions of the manufacturer.