GSM5116216: Urine_Pb_1mo_rep2; Rattus norvegicus; miRNA-Seq
Total RNA was isolated from urine samples using the Urine Cell-Free Circulating RNA Purification Mini Kit (Cat. 56900, Norgen Biotek, Thorold, ON, Canada) following manufacturer's protocol. Briefly, 2 ml of cell-free urine was mixed with Binding Solution K and transferred to spin column. RNA within the sample was bound to the spin column while the urine was discarded as flow-through. Following application of Lysis Buffer A, the flow-through was precipitated with isopropanol and subjected to repeat centrifugation through the column to allow maximum RNA binding. The column was then washed twice with Wash Solution A, centrifuged to remove left over alcohol, and transferred to a new RNAse-free tube for elution. The concentration of total RNA was assessed with the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA). Library preparation and sequencing was performed by Norgen Biotek. Briefly, library preparation was done using the Norgen Biotek Small RNA Library Prep Kit (Cat. 63600). Sequencing was performed using the NextSeq 500/550 High Output Kit v2 (51 Cycles using a 75-Cycle Kit) on the Illumina NextSeq 500 platform.