title: GSM3671270: Upf1_1_PARCLIP; Saccharomyces cerevisiae; RIP-Seq
library_construction_protocol: The cells were lysed by bead beating and IP was performed with rabbit IgG-conjugated protein G magnetic Dynabeads (Invitrogen). The cross-linked RNA was partially digested with RNase T1 (Thermo Scientific) and the IP was controlled by performing a western blot analysis of a part of the sample using the Peroxidase Anti-Peroxidase (PAP; Sigma) antibody. Following adapter ligation, RNA was recovered by digestion with proteinase K (New England Biolabs) and a subsequent acidic phenol/chloroform extraction. Reverse transcription was performed with SuperScript III RTase (Invitrogen) and was followed by PCR amplification using the NEXTflex barcode primer kit (Bio Scientific). The generated cDNA was purified, size selected, and quantified using TapeStation (Agilent Technologies). The resulting samples were then PCR-amplified and sequenced on an Illumina HiSeq 1500, HiSeq2500 or NextSeq550 system.
attributes.tag: GEO Accession
attributes.value: GSM3671270