GSM2331470: TAA_Low_3day_45; Rattus norvegicus; miRNA-Seq
Total RNA was isolated from approximately 15 mg liver sections using miRVanaTM isolation kit (Ambion® ThermoFisher Scientific, Grand Island, NY, USA) according to manufacturer's protocol. The yield of the extracted RNA was determined spectrophotometrically by measuring the optical density at 260 nm (Nanodrop-1000, Thermo Scientific, Wilmington, DE, USA). The quality of RNA samples was characterized on an Agilent BioAnalyzer (Agilent Technologies, Santa, Clara, CA, USA) using an RNA6000 Nano Chip (Agilent Technologies). RNA integrity numbers (RINs) greater than 8.0 were used for NGS analysis with an average of 9.2 for all samples. 1 µg of extracted total RNA was used for miRNA-seq library preparation using Illumina TruSeq small RNA sample preparation kit. Each RNA sample was processed individually by adding special bar-coded adaptors and reverse transcriptase and amplified. The quantity and quality of each library was measured by Picogreen DNA quantification and Bioanalyzer high sensitive Pico chips (Agilent Technologies). Forty eight barcoded libraries were then pooled by mixing equi-molar of DNA. The pooled libraries were then loaded onto 6% high resolution PAGE gel for separation of miRNA from other RNAs. The bands with the molecular weight between 145-160 bp were excised into recovery tubes for gel purification. The recovered barcoded libraries were then sequenced by the Illumina HiSeq2000 platform with single end base pair reads.
GEO Accession
GSM2331470