GSM6883037: GM1-Neuron2mo-WT; Homo sapiens; RIP-Seq
Neurons were crosslinked with 254-nm UV-C followed by cell lysis and partial digestion of RNAs. FMRP-bound RNAs were isolated by immunoprecipitation, dephosphorylated, and ligated to a dye-labeled 3′ adapter. RNA samples were then resolved using SDS-PAGE and transferred to nitrocellulose membranes. FMRP-bound RNAs were visualized and size selected by cutting the membranes. Size-selected RNAs were purified by Proteinase K digestion and ethanol precipitation. The purified RNAs were converted to cDNAs followed by circularization of the cDNAs. cDNA libraries were constructed from the cDNAs by PCR amplification.