GSM4826757: Yil161w_input_rep2; Saccharomyces cerevisiae; OTHER
Cell lysates were treated with RNase, and ribosomes were sedimented through 1 M sucrose cushion. RNA from the ribosome fraction was extraced using TRIzol reagent (Thermo Fisher Scientific) following manufacturer's instructions. Libraries were prepared for sequencing using a customized protocol from Ingolia et al., 2012. In brief, ribosome-protected mRNA fragments were purified and ligated to a single-stranded linker that serves as a priming site for reverse transcription. The first-strand reverse transcription products are circularized and PCR-amplified to generate a deep-sequencing library.