GSM2147543: CB3; Homo sapiens; miRNA-Seq
Total RNA was extracted using the mirVana miRNA Isolation Kit from ThermoFisher Scientific Inc. (Waltham, MA, USA) according to the recommended procedure. The quantity of RNA yield was determined with NanoDrop from ThermoFisher Scientific Inc. (Waltham, MA, USA), and the quality was assessed using the RNA 6000 Nano Kit with Bioanalyzer 2100 from Agilent Technologies (Santa Clara, CA, USA). The TruSeq Small RNA Sample Prep kit from Illumina (San Diego, CA, USA) was used to generate the small RNA sequencing library. Briefly, 1 µg of total RNA was ligated using the manufacturer supplied RNA 3’ and RNA 5’ adapters. To produce cDNA constructs, the ligated small RNA was subjected to RT-PCR with a sample specific index sequence followed by gel purification. Amplified miRNAs were enriched and validated with the Agilent Bioanalyzer 2100 using High Sensitivity DNA chips. Using a MiSeq Reagent Kit v2 with 50 cycles, the validated small RNA sequencing libraries were normalized, denatured, and loaded to the Illumina MiSeq sequencer.
GEO Accession
GSM2147543