Automated Ultra-Long DNA and RNA Extraction for Long-read Sequencing Applications

Summary Claremont BioSolutions, LLC (CBIO) The ability to sequence and map long stretches of DNA and RNA using third generation sequencing technologies is set to transform the field of genomics and allow researchers and clinicians unparalleled insight into the complexities of our genome and diseases. However, the move to these platforms presents new hurdles in long DNA and RNA sample preparation. Most conventional spin column sample preparation methods shear DNA during purification, and although agarose plug digestion methods can isolate ultra-long DNA from samples, the protocols can be difficult, time consuming and highly variable. While newer technologies have emerged that allow for more rapid extraction of long-DNA, these kits are either limited in use to easy-to-lyse sample types (cells and blood) or require extra equipment and long processing times, which significantly increases the cost. At Claremont BioSolutions (CBio), we are developing a novel automated extraction technology that will incorporate lysis, nucleic acid binding and isolation. The technology will allow users to add and sample, hit “GO” and in 30 minutes collect DNA or RNA. In Aim 1, we will integrate our fast lysis and isolation technology into an automated cartridge to provide reproducible high quality, ultra-long DNA or RNA extraction. In Aim 2, we will validate automated ultra-long DNA extraction using easy and complex sample types, compare our technology to HMW DNA competitor's kits, and demonstrate improved long-read sequencing using two third generation technologies. In Aim 3 we will test automated RNA extraction and downstream native RNA sequencing. In Aim 4 we will perform pre-market analysis at off-site alpha and beta testing facilities to generate end-user feedback and lockdown the design. The proposed system would simplify workflow, lower cost, and reduce variability.