GSM9092172: HEK293T-EF1α-mCherry (WT), 2; Homo sapiens; OTHER
ribosome profiling was conducted following the protocol for GenSeq® Ribo Profile Kit (GenSeq, Inc). Briefly, cycloheximide-treated cells were lysed with lysis buffer and digested with nuclease. Ribosome protected fragments (RPFs) were obtained through size exclusion chromatography, size selection by polyacrylamide gel-electrophoresis (PAGE), and rRNA removal. The purified RNA fragments were then end-repaired, ligated with 3' adapters and reverse-transcribed into cDNA, which was subjected to PAGE-purification, circularization, and PCR-amplification.