GSM5746664: L_BWM_rep05_batchA; Rattus norvegicus; RNA-Seq
Livers were removed and frozen in Eppendorf tubes using liquid nitrogen immediately. Tissue was homogenized using QIAGEN Tissue Lyser II (85300; QIAGEN) and further purified using Proteinase K (PK) Solution (MC5005; Promega). Prior to extraction, RNA integrity and concentration were tested using NanoDrop™ 2000c spectrophotometer (Thermo Fisher Scientific). In the following step, mRNA was extracted using Promega's “Maxwell SimplyRNA Tissue Kit” (AS1340). RNA quality was checked using RNA 6000 Nano kits on a 2100 Bioanalyzer (Agilent Technologies). Afterwards, cDNA libraries were prepared from total RNA with oligo-dT capture beads for poly(A)-mRNA enrichment using the TruSeq Stranded mRNA Library Preparation Kit (Illumina) according to manufacturer's instructions. After 15 cycles of PCR amplification, barcoded cDNA libraries were quantified via Qubit 2.0 Fluorometer using dsDNA HS Assay Kits (ThermoFisher). Quality and size distribution (average size: 290 bp) was checked with a 2100 Bioanalyzer using DNA 1000 kits (Agilent).