GSM7017408: 1 mg furan 21d rep 4; Mus musculus; miRNA-Seq
Total RNA was extracted from frozen liver tissue (n=4-5/treatment) using a ceramic bead-based homogenization combining an initial RNAzol RT® (MRC, Cincinnati, OH) extraction with subsequent purification using the RNeasy MinElute kit (Qiagen, Germantown, MD).  Clean-up used the modifications for RNA binding indicated in the MinElute kit protocol supplement to retain the small RNA fractions.  Total RNA from whole blood (n= 4-5/treatment) was isolated using the mirVANA RNA isolation kit (ThermoFisher Scientific, Waltham, MA) and from in vitro samples (n=2/treatment) using the miRCURY RNA isolation kit (Exiqon Inc., Woburn, MA).  RNA was quantified by Nanodrop spectrophotometer (NanoDrop Technologies, Wilmington, DE) and/or Qubit fluorometric assay (ThermoFisher Scientific).  All RNA exhibited high purity with A260/280 ratios >2.0. To create sequencing libraries, the TruSeq Small RNA Library Prep Kit was utilized following manufacturer's instructions. Quality and size range of prepped libraries were check using Agilent Bioanalyzer before loading on flow cell.