A custom library construction method for super-resolution ribosome profiling in Arabidopsis.

ABSTRACT:

Background

Ribosome profiling, also known as Ribo-seq, is a powerful technique to study genome-wide mRNA translation. It reveals the precise positions and quantification of ribosomes on mRNAs through deep sequencing of ribosome footprints. We previously optimized the resolution of this technique in plants. However, several key reagents in our original method have been discontinued, and thus, there is an urgent need to establish an alternative protocol.

Results

Here we describe a step-by-step protocol that combines our optimized ribosome footprinting in plants with available custom library construction methods established in yeast and bacteria. We tested this protocol in 7-day-old Arabidopsis seedlings and evaluated the quality of the sequencing data regarding ribosome footpr

SUBMITTER: Wu HL 

PROVIDER: S-EPMC9531494 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature