GSM6892033: Polysome-Seq WT2, replicate2; Mus musculus; OTHER
Cell lysates were centrifuged at 18,000 x g at 4 ℃ for 15 min and the supernatant was digested with RNase Ⅰ. Ribosome-protected RNA fragments (RPF) were obtained using MicroSpin S-400 column resin; the was column enriched for ribosome-mRNA complexes and the targeted RNA fragments were purified by PAGE. The purified RNA fragments were end-repaired and A-tailed, and 5' and 3' joints were directly added to the ends. RNA reverse transcriptase was used to synthesize cDNA, and additional steps of PCR enrichment and PAGE-based screening of target fragments were used to obtain a library containing target fragments. The library was verified by quality control screening and sequencing. Ribosome-seq