RNA sequencing of nuclease dead PET127 conditions

PET127, is thought to be a mitochondrial RNA exonuclease and loss of PET127 is thought to cause accumulation of untrimmed RNA in the mitochondria. Here we show RNA sequencing of Saccharomyces cerevisiae in four PET127 conditions: PET127 wild-type, pet127∆, pet127-nd (nuclease dead)/pet127∆, and pet127-nd/PET127. The resulting sequences show that regions of mitochondrial RNA accumulate in the pet127∆ background, and the RNA sequencing profile of PET127 containing disruptive mutations in the conserved exonuclease active site mimics that of pet127∆. 

Methods
RNA was isolated from 4.4 x 107 log phase cells using acid phenol and purified using RNeasy kit (Qiagen) with on column DNAse treatment (Qiagen). Isolated total yeast RNA were processed using Ribozero rRNA removal for Yeast (Illumina). Sequencing was performed on NextSeq500 with 75 + 75 bases pair-end run with 6 + 6 nucleotide indexes.