GSM4790534: Dioxane 5000mg/mL rep3; Mus musculus; RNA-Seq
mRNA (≈500 ng total) was purified with oligo-dT beads and sheared by incubation at 94°C (first-strand synthesis with random primers). Second strand synthesis was performed using dUTP to generate strand-specific sequencing libraries. The cDNA library was then end-repaired, A-tailed adapters were ligated, and second-strand digestion was performed using uracil-DNA-glycosylase. Indexed libraries (that met appropriate cut-offs for both) were quantified by qRT-PCR using a commercially-available kit (KAPA Biosystems), and insert size distribution was determined using a LabChip GX or Agilent Bioanalyzer. Samples with a yield ≥0.5 ng/µL were used for sequencing. RNA libraries were prepared for sequencing using standard Illumina protocols