GSM2992209: Stau2 E13.5 Rep 1; Mus musculus; RIP-Seq
RIPs were performed with the MAGNA-RIP kit (Millipore, 17-700). For each RIP, 15 μl of Protein A/G beads were blocked with 2% UltraPure BSA (ThermoFisher, AM2616) at room temperature (RT) for 1-2 hours (hrs) before overnight incubation with 5-10 ug of either mouse anti-STAU2 (Abcam ab60724) or mouse anti-T7 (Novagen/Millipore 69522-3) monoclonal antibodies. For generation of the RIP-seq dataset, STAU2 RIP results were compared directly to input samples. After generation of the RIP-Seq dataset, a small number of select targets were compared with results from negative control T7-RIPs, using qPCR rather than direct sequencing. RNA was purified using TRIzol (ThermoFisher) and RNeasy Micro (Qiagen, 74004) or RNA Clean and Concentrator (Zymo, R1014) Kits, with DNase treatment. For RIPs performed on whole cortical lysate, pooled cortical tissue from a whole litter of embryos was used as starting material for each independent experiment (i.e. biological replicate). For RIPs performed on TBR2+ GFP-high cells, cell fractions obtained from two separate FACS collections were pooled and used as starting material for each independent experiment (i.e. biological replicate). Libraries for RNA-seq were prepared using 'ScriptSeq™ Complete Kit (Human/Mouse/Rat)-Low Input from Epicentre/Illumina using manufacturer's instructions.
GEO Accession
GSM2992209