GSM2483279: SUP-26_CLIP_repA; Caenorhabditis elegans; OTHER
Pellets were thawed in cold homogenization buffer [100mM NaCl, 25mM HEPES pH=7.5, 250µM EDTA, 2mM DTT, 25U/ml RNAse inhibitor (ThermoFisher RiboLock), protease inhibitors (Complete Mini, Roche)] at 4°C. Samples were sonicated on ice at 30W using 20s pulses with 60s rest intervals (Branson SLPe). Sonication continued until pellet was nearly completely homogenized (such that only small volume pellet, ~50µl, was visible after the sample settles). Samples were centrifuged at 16000rcf at 4°C for 15min, and supernatant was moved to a fresh tube. Protein was quantified, and concentration was adjusted to 3mg/ml in homogenization buffer. CLIP libraries were prepared as in Zisoulis et al., 2011, with cloning steps modified from Gu et al., 2011. See supplemental methods in McJunkin & Ambros, "A microRNA family exerts maternal control on sex determination in C. elegans".
GEO Accession
GSM2483279