title: GSM5683521: 4h_tg_5_10769; Mus musculus; RNA-Seq
library_construction_protocol: Both cohort of mice were sacrificed at ZT7.5 and liver samples were extracted and snapped frozen in liquid nitrogen. Total murine liver RNA was isolated using the mirVana Isolation Kit (Ambion) in accordance to the manufacturer's instructions. Murine liver RNA concentration was measured by Nanodrop ND-100 (Peqlab Biotechnology, France), and diluted to a final concentration of 200 – 250 ng/mL. RNA integrity number (RIN) was analyzed with the Agilent RNA 6000 Nano Kit (5067-1511, Agilent Technologies) and the Agilent 2100 Bioanalyzer. All samples displayed a RIN of >8. RNA libraries were prepared with 1 µg of total RNA using the Illumina® TruSeq® mRNA stranded sample preparation Kit. After poly-A selection (using poly-T oligo-attached magnetic beads), mRNA was purified and fragmented using divalent cations under elevated temperatures. The RNA fragments underwent reverse transcription using random primers. This is followed by second-strand cDNA synthesis with DNA Polymerase I and RNase H. After end repair and A-tailing, indexing adapters were ligated. The products were then purified and amplified (14 PCR cycles) to create the final cDNA libraries. After library validation and quantification (Agilent tape station), equimolar amounts of the library were pooled. The pool was quantified by using the Peqlab KAPA Library Quantification Kit and the Applied Biosystems 7900HT Sequence Detection System.