Next-generation sequencing of double stranded RNA is dramatically improved by treatment with an inexpensive reagent

Double stranded RNA (dsRNA) serves as the genome of important viruses and is a key component of RNA interference-based immunity in eukaryotes. DMSO has been used to denature dsRNA prior to reverse transcription stage to improve RT-PCR and Sanger sequencing. We systematically tested the utility of DMSO to improve sequencing yield of a dsRNA virus (Φ6) in a short-read next generation sequencing platform. We found that DMSO treatment improved sequencing read recovery by over two orders of magnitude, even when RNA and cDNA concentrations were below the limit of detection. Furthermore, we provide evidence that this treatment does not adversely affect recovery of reads from single-stranded RNA viral genomes (influenza A/California/07/2009). We suggest that 50% DMSO treatment is used prior to cDNA synthesis when samples of interest are or may contain dsRNA.These libraries were prepared from ssRNA genomes, treated with varying concentrations of DMSO before cDNA synthesis.