ferret_371_day0
Following the manufactures protocols, where available 100 ng of total RNA was used as input material in to the QIAseq FastSelect rRNA (HMR) (Qiagen) protocol to remove cytoplasmic and mitochondrial rRNA with a fragmentation time of 7 minutes. Subsequently the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs) was used to generate the RNA libraries. Following 13 cycles of amplification the libraries were purified using Ampure XP beads. Each library was quantified using Qubit and the size distribution assessed using the Fragment analyser. These final libraries were pooled in equimolar amounts using the Qubit and Fragment analyser data. The quantity and quality of the pool was assessed by the Bioanalyzer and subsequently by qPCR using the Illumina Library Quantification Kit from Kapa on a Roche Light Cycler LC480II according to manufacturer's instructions.