title: GSM2940087: eCLIP.untagged_1.low_RNase; Saccharomyces cerevisiae; RIP-Seq
library_construction_protocol: Cells were lysed by vortexing with glass beads in lysis buffer (50 mM Tris-HCl pH 7.4, 100 mM NaCl, 1% NP-40, 0.1% SDS, 0.5% sodium deoxycholate, 1:200 Protease Inhibitor Cocktail III, 0.44U/mL murine RNase inhibitor). Extracts were clarified at 10,000 x g for 5 minutes. Lysate was digested with 1:20,000,000 RNase I and immunoprecipitated with anti-c-Myc-conjugated beads. A 3′ adapter was ligated and the RNPs separated on a denaturing gel. 55-130 kDa species were size-selected and treated with proteinase K. RNAs were reverse transcribed. A second adapter was ligated to the 3′ end of first-strand cDNA. cDNAs were then amplified by PCR.
attributes.tag: GEO Accession
attributes.value: GSM2940087