Instrument: Illumina NovaSeq 6000
Strategy: OTHER
Source: TRANSCRIPTOMIC
Selection: other
Layout: SINGLE
Construction protocol: Sample was treated with specific lysis buffer with cycloheximide (50mg/mL) to acquire the lysate. Cells were treated with specific lysis buffer of cycloheximide to stop translation. Then, cell lysate was treated with unspecific endoribonuclease RNase to digest the free RNAs other than ribosome-protected fragments (RPFs). The RNA samples were then treated with rRNA depletion kit to remove rRNA contamination. Small RNA (< 200 bp) was resolved by PAGE, and the RPFs bands (~30nt) were cut for RNA extraction, cDNA transcription, and PCR amplification.