GSM2349426: dCLIP Cbx7 rep 3; Mus musculus; RIP-Seq
UV-cross-linked cells, expressing BirA biotinylase and Biotin-tagged Cbx7 or mock vector, were isolated and lysed. Lysates were DNase-treated and RNAse-treated. Cleared lysates were incubated 2hrs with Streptavidin beads. After stringent washing and 2nd DNase treatment, CLIP-tags on beads were radiolabelled with [γ-32P]ATP, run on Bis-Tris SDS-PAGE in MOPS buffer, and transferred to nitrocellulose membrane. Membrane fragments containing CLIP signal and corresponding positions on control lanes were excised, and RNA was eluted by proteinase K, followed by Trizol extraction and ethanol precipitation. Multiplex Compatible NEBNext Small RNA Library Prep Set for Illumina (NEB). Illumina barcode #12, CTTGTA. Paired-end 50bp reads