Automated RNA Purification from Salmonid Organs Using a Novel Paramagnetic Particle Technology

RNA viruses, including viral hemorrhagic septicemia virus, Salmonid alphaviruses (SAV) and infectious salmon anemia virus (ISAV), pose a significant threat to Atlantic salmon populations. Molecular techniques offer the means of monitoring for these viruses in fish farms as well as wild salmon populations. To facilitate rapid, reliable testing, we explore the use of magnetic particle-based purification technologies for automated extraction of RNA from salmon and trout organs. Two automated methods were examined, one is the Maxwell® 16 Instrument with the LEV simplyRNA Tissue Kit and the second is a custom method and reagents that were created to adapt the purification chemistry to a 96-well format for laboratory liquid handlers such as the Biomek 4000 Laboratory Automation Workstation. Using optimized reagents and protocols, we were able to extract more than 4.5µg of RNA from salmon brain, gill, and heart tissues, and more than 20µg of RNA from salmon kidney, liver and spleen tissues in both low throughput and high throughput automated processing. The RNA was of high integrity and purity and performed well in subsequent RT-qPCR assays. Two transcripts (β-actin and EIF1α) were monitored in all tissues and showed linear detectability across 5 orders of magnitude dilutions of the RNA samples. These data support the use of the simplyRNA reagents on the Maxwell® 16 Instrument and customized for 96 well processing allowing automated purification of salmon organ RNA to streamline monitoring of salmon populations.