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Unless other concentrations are specified, 0.75 nM of pre-diluted AGO–miRNA complex was pre-incubated with approximately 0.25 nM target RNAs in co-IP buffer (18 mM HEPES pH 7.4, 150 mM KOAc, 2 mM Mg(OAc) 2 , 5% (v/v) glycerol, 0.01% (v/v) IGEPAL CA-630 and 0.5 mM BME) supplemented with blocking reagents (0.1 mg ml −1 recombinant albumin and 0.01 mg ml −1 yeast tRNA) at 37 °C for 30–60 min