GSM2653253: RIP8 IP CPEB1; Homo sapiens; RIP-Seq
RNP-complexes were crosslinked with 0.5% PFA for 5 min, then CPEB1/4 were immunoprecipitated with anti-CPEB1/4 specific antibodies and associated-mRNAs were extracted with phenol-chlorophorm. Inputs and IPs samples from a RIP experiment were quantified by Qubit RNA HS (Life Technologies) when it was possible, and fragment size distribution was checked by Bioanalyzer Pico Chip (Agilent). Library generation was done with the kit NEBNext Ultra RNA Library Prep Kit (New England Biolabs), following manufacturer´s instructions and 14-15 cycles of library amplification. Previous Poly A purification was only performance on Input samples in only one step of purification. Size selection of the libraries and purification were done by magnetic beads (Agencourt AMPure XP, Beckman Coulter). Finally, library quantification was performance with Qubit DNA HS (Life Technologies) and fragment size distribution with Bioanalyzer DNA HS (Agilent)
GEO Accession
GSM2653253