Diversity of phytoviral RNA U-tailing among ss(+) RNA phytoviruses

Using 3' RACE-seq method, we analyzed global 3’ terminal RNA uridylation profiles for representatives of the main families of positive single-stranded RNA phytoviruses. 3’ RACE-seq method was used to measure poly(A) tail sizes and analyze uridylation levels. This protocol is based on the ligation of a primer at the 3’ end of RNA, and the subsequent targeted amplification by PCR of amplicons suitable for Illumina sequencing.

本研究采用3'末端快速扩增测序（3' RACE-seq）技术，对主要科属的正链单链RNA植物病毒代表样本的全局3'端RNA尿苷化谱进行了分析。该技术可同时用于测定poly(A)尾长度，并分析RNA的尿苷化水平。本实验方案基于在RNA的3'末端连接引物，随后通过PCR对适配Illumina测序的扩增子进行靶向扩增。