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Interest in RNA-protein interactions is booming as we begin to appreciate the role of RNA, not just in well-established processes such as transcription, splicing and translation, but also in newer fields such as RNA interference and gene regulation by non-coding RNAs. RIP is an antibody-based technique used to map RNA–protein interactions in vivo by immunoprecipitating the RNA binding protein of interest together with its associated RNA and allows identification of bound transcripts. RIP precipitates a specific RNA binding protein (RBP) and associated RNA (mRNAs, non-coding RNAs, viral RNAs) that can be detected by real-time PCR, microarrays or e.g. sequencing. Here is a RIP protocol adapted from Khalila et al, 2009, Hendrickson et al, 2009, Hendrickson et al, 2008 and from Rinn et al, 2007.
Nuclear isolation buffer