GSM6601660: Sample 3-IP; Mus musculus; RIP-Seq
Total RNA was subjected to immunoprecipitation with the GenSeq® m5C-IP Kit (GenSeq Inc.) by following the manufacturer's instructions. Briefly, RNA was randomly fragmented by RNA Fragmentation Reagents. Protein A/G beads were coupled to the m5C antibody by rotating at room temperature for 1 h. The RNA fragments were incubated with the bead-linked antibodies and rotated at 4°C for 4 h. After incubation, the RNA/antibody complexes were washed for several times, and then, captured RNA was eluted from the complexes and purified. Both the input samples without immunoprecipitation and the m5C IP samples were used for RNA-seq library generation with NEBNext® Ultra II Directional RNA Library Prep Kit.  Library sequencing was performed on an illumina Hiseq instrument with 150bp paired-end reads.