Protocol to measure ribosome density along mRNA transcripts of Arabidopsis thaliana tissues using Ribo-seq

利用Ribo-seq技术测量拟南芥组织中mRNA转录本核糖体密度的实验方案

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作者:Isabel Cristina Vélez-Bermúdez ,Shu-Jen Chou ,Ai-Ping Chen ,Wen-Dar Lin ,Wolfgang Schmidt

Abstract

Ribosome profiling (Ribo-seq) measures ribosome density along messenger RNA (mRNA) transcripts and is used to estimate the "translational fitness" of a given mRNA in response to environmental or developmental cues with high resolution. Here, we describe a protocol for Ribo-seq in plants adapted for the model plant Arabidopsis thaliana. We describe steps for lysis and nucleolytic digestion and ribosome footprinting. We then detail library construction, sequencing, and data analysis. Keywords: Genomics; Model Organisms; Molecular Biology; Plant Sciences; RNAseq.

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