I-Corps: Amplification and Next-Generation Sequencing Library Preparation Using Picogram Quantities of Total RNA/DNA

The broader impact/commercial potential of this I-Corps project is the development of a technology to sequence DNA/RNA from biological materials in small quantities. High-throughput RNA/DNA sequencing, called Next-Generation Sequencing (NGS), is a key technology in healthcare, medical research, and life sciences. It is used in the diagnosis of emerging infectious diseases, biopsies, pre-natal diagnostics, organ transplant, forensics, archaeology, and also basic biological and medical research. However, sequencing of the biomaterials available in small quantities is challenging and most precious biomaterials are usually limited in availability. Single cell genomics and spatial genomics, which also involve small quantity samples, are predicted to be important technologies in the future that will drive the basic understanding of diseases, diagnostics, and therapeutics. The proposed technology may broaden the use of NGS and serve as an enabling technology for this rapidly growing area.This I-Corps project is based on the development of technology to generate DNA/RNA sequence information using Next-Generation Sequencing (NGS) from small quantities of biological materials. Market research suggests that about ~40% of laboratories believe that the biggest bottleneck in NGS is in “library preparation methods.” The proposed NGS library preparation technology may increase the scope of NGS applications. The proposed technology has been shown to be 2-5 times more efficient and generates both quantity and quality data using a lower quantity of template (1-1/100th) compared with existing methods. In addition, the proposed technology may be used for sequencing of small-RNAs using sub-nanogram (picogram) quantity of template, which is not possible using existing technologies. This use of smaller sample sizes may allow NGS to be used with new biological materials and be more accurate, reliable, and informative, which may improve diagnostic and other results.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.