Analysis of the Human Mitochondrial Transcriptome Using Directional Deep Sequencing and Parallel Analysis of RNA Ends

RNA sequencing using next-generation technologies provides comprehensive coverage of transcriptomes at a much greater depth than conventional transcriptomic methods. The human mitochondrial genome is relatively small, and sequencing its transcriptome provides a valuable method to investigate changes in RNA metabolism in great detail. Here we describe two methods that use next-generation technologies to investigate mitochondrial RNAs. Directional RNA sequencing enables the analyses of RNA abundance from each strand of the mitochondrial DNA. Parallel analysis of RNA ends enables the analyses of processing of mitochondrial transcripts, their termini, and annotation of any new transcripts.