High density analysis of the tailing and trimming patterns of RISC-cleaved MYB33 and SPL13 transcripts in Arabidopsis heso1 and urt1 mutants

Using 3' RACEseq method, we compared the tailing and nibbling patterns of RISC-cleaved MYB33 and SPL13 transcripts between wild-type plants and mutant plants depleted for the terminal uridylyltransferases (TUTases) HESO1 and URT1. Our data reveal a major and minor role for HESO and URT1, respectively, in the uridylation of RISC-cleaved transcripts. 3’ RACE-seq method was used to analyze the 3’ ends of 5’ MYB33 and SPL13 RISC-cleaved transcripts for urt1, heso1 and heso1 urt1 mutants. 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.