Normalized Ribo-Seq for Quantifying Absolute Global and Specific Changes in Translation.

ABSTRACT: Ribosome profiling (Ribo-Seq) is a highly sensitive method to quantify ribosome occupancies along individual mRNAs on a genome-wide scale. Hereby, ribosome-protected fragments (= footprints) are generated by nuclease digestion, isolated, and sequenced together with the corresponding randomly fragmented input samples, to determine ribosome densities (RD). For library preparation, equal amounts of total RNA are used. Subsequently, all transcript fragments are subjected to linker ligation, cDNA synthesis, and PCR amplification. Importantly, the number of reads obtained for every transcript in input and footprint samples during sequencing depends on sequencing depth and library size, as well as the relative abundance of the transcript in the sample. However, the information pertaining to the a

SUBMITTER: Hoerth K 

PROVIDER: S-EPMC8899559 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature