GSM8178890: liver, PXR-CAR-CYP3A4/3A7_ MI-883_5; Mus musculus; RNA-Seq
Total RNA from liver samples has been isolated using TRIZOL®. Total RNA was purified using Agencourt RNAClean XP Beads (Beckman Coulter, Inc.) and then the diluted RNA was quantified using Qubit 2.0 Fluorometer with Qubit RNA BR Assay Kit (Invitrogen) for precise measurement. The integrity of purified RNA was evaluated based on RIN (>7) acquired using the 2100 Bioanalyzer System with Total RNA Nano Chip (Agilent Technologies) according to the manufacturer's protocol. The input amount of total RNA used in the libraries was 520 ng. Libraries were constructed with NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs Inc., Ipswich, MA, USA). The whole procedure was performed according to the protocol suggested by the manufacturer. The samples were multiplexed using suitable molecular barcodes by using the NEBNext Multiplex Oligos from Illumina. Adaptor enrichment was performed by using eight cycles of PCR. The concentration of molecular libraries was measured using Qubit 2.0 Fluorometer with Qubit dsDNA HS Assay Kit (Invitrogen).