GSM2150443: 285 Heart 03; Rattus norvegicus; miRNA-Seq
Frozen tissues (heart, lung, liver, and kidney) stored at -80 °C in cryovials were placed on dry ice, cut into aliquots with a sterile scalpel on a pre-chilled titanium block, and placed in new tubes pre-chilled on dry ice. While working on dry ice, whole tissues (~25 mg) from the lung, liver, and kidney were cut from each sample and placed in 700 mL of QIAzol Lysis Reagent. Samples were homogenized with a TissueLyzer LT (Qiagen, Valencia, CA) for 5 min at 25 Hz twice, allowed to sit at room temperature for 5 min, and then placed at -80 °C. We homogenized heart tissue with a 6750 Freezer Mill with three sample microvials (Spex SamplePrep, Metuchen, NJ). RNA was isolated using the miRNeasy 96 kit according to the manufacturer’s instructions. The quality and quantity of RNA samples were evaluated with a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA) using the Agilent RNA 6000 Nano Reagents and a multi-well Nanodrop 8000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA). Small RNA libraries were constructed using the TruSeq small RNA library preparation kit (Illumina, San Diego, CA) according to the manufacturer’s instructions.
GEO Accession
GSM2150443