Sequencing the RNA in a biological sample can unlock a wealth of information, including the identity of bacteria and viruses, the nuances of alternative splicing or the transcriptional state of organisms. However, current methods have limitations due to short read lengths and reverse transcription or amplification biases. Here researchers at Oxford Nanopore Technologies demonstrate nanopore direct RNA-seq, a highly parallel, real-time, single-molecule method that circumvents reverse transcription or amplification steps. This method yields full-length, strand-specific RNA sequences and enables the direct detection of nucleotide analogs in RNA.
(a) Library preparation method for direct RNA-seq. (b) Representative raw data ‘squiggle’ resulting from translocation of a single transcript through a pore in the MinION array. (c) Alignment of a typical Saccharomyces cerevisiae S228C read to the reference transcriptome.