GSM7949404: NC [Ribo-seq]; Mus musculus; OTHER
Ribosome profilng service was provided by CloudSeq Inc.(Shanghai, China) by following the manual for the GenSeq® Ribo Profile Kit (GenSeq,Inc.). Briefly, the cycloheximide-treated cells were lysed with lysis buffer and digested with nuclease. Digested samples were purified with size exclusion chromatography to obtain ribosome footprints. Ribosome protected fragments were size-selected with polyacrylamide gel electrophoresis (PAGE), and subjected to rRNA removal. Then, purified RNA fragments were end-repaired, ligated with 3' adapters and reverse-transcribed to cDNA. The cDNA was PAGE gel purified, circularized, and amplified by PCR. The resulting libraries were purifed and sequenced in a NovaSeq platform (Illumina).