GSM4670970: TAT-02: Wild type Male_HFHC_Liver; Mus musculus; RNA-Seq
RNA was isolated from snap frozen tissue using the miRNeasy Mini Kit (QIAGEN) according to manufacturer's instruction with an elution volume of 45 µl. The library preparation was done using TruSeq® stranded total RNA sample preparation kit with rRNA depletion (Illumina inc). Briefly, the starting material (100 ng) of total RNA was rRNA depleted using biotinylated, target-specific oligos combined with Ribo-Zero rRNA removal beads. The isolated RNA was subsequently fragmented using enzymatic fragmentation. Then first strand synthesis and second strand synthesis were performed and the double stranded cDNA was purified (AMPure XP, Beckman Coulter). The cDNA was end repaired, 3' adenylated and Illumina sequencing adaptors ligated onto the fragment ends, and the library was purified (AMPure XP). The RNA stranded libraries were pre-amplified with PCR and purified (AMPure XP). The libraries' size distribution was validated and quality inspected on a 4200 TapeStation (Agilent Technologies). High quality libraries were pooled in equimolar concentrations based on the Bioanalyzer automated electrophoresis system (Agilent Technologies). The library pool(s) were quantified using qPCR and optimal concentration of the library pool used to generate the clusters on the surface of a flow cell before sequencing on a NextSeq500 instrument (2x75 cycles) according to the manufacturer instructions (Illumina Inc.).
GEO Accession
GSM4670970