GSM5708603: RiboSeq_WT3; Mus musculus; RNA-Seq
For the RiboSeq experiment, ribosome protected fragments (RPFs) were generated using RNAse I treatment and isolated using MicroSpin S-400 columns (Cytiva). For the RNA-Seq experiment, total RNA was isolated from the lysate using TRIzol. For the RiboSeq experiment, purified RPFs were used for the library preparation using the NEXTFLEX small-RNAseq V3 kit (Perkin-Elmer). For the RNA-Seq experiment, libraries were prepared using NEBNext® Poly(A) mRNA Magnetic Isolation Module (ref. e7490) and NEBNext® Ultra II Directional RNA Library Prep Kit for Illumina (24 reactions ref. e7760 or 96 reactions ref. e7765) according to the manufacturer's protocol, to convert total RNA into a library of template molecules of known strand origin and suitable for subsequent cluster generation and DNA sequencing.