Ribo-Seq of gallus: myoblast
In this study, the longest transcript of each protein-coding gene was first screened, and non-canonical ORFs (uORF, dORF, lincORF) with a length of 60450 nt were identified in non-coding regions (5UTR, 3UTR, and lincRNA). After RNA removal and alignment, the RPKM method was used to quantify the translation level of ORFs. Furthermore, the ribosome release score (RRS) and ORF score were combined to evaluate the coding potential, and the 95th percentile value of known coding genes was used as the threshold to screen highly reliable translated ORFs. Differentially translated ORFs (DT ORFs) were identified by edgeR (|FC|2, FDR<0.05). Meanwhile, the CPAT tool and Pfam domain analysis were used to assist in verifying the coding potential of non-canonical ORFs.