Research Project
9681228
Summary Claremont BioSolutions, LLC (CBIO) The ability to sequence and map long stretches of DNA 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 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 platforms are limited in starting sample type to fresh blood or tissue culture cells and exclude use of most clinically relevant frozen blood and solid tissue samples. At Claremont BioSolutions (CBIO), we are developing a novel extraction technology that utilizes a proprietary high capacity nucleic acid binding matrix, which is loosely packed in a low pressure fast flow column. Combined with CBIO's gentle upfront lysis method, the technology will be compatible with any starting tissue sample type, including frozen blood and solid tissue, and can be used to isolate ultra-long DNA in <30 minutes. In Aim 1, we will demonstrate CBIO's fast-flow isolation of high quantity, high quality, ultra-long DNA (>250 kilobases) from fresh and frozen blood and tissue samples and benchmark against existing extraction methods. In Aim 2, we will validate isolated ultra-long DNA compatibility with two third generation long-read sequencing and mapping technologies and demonstrate improved long-read length over existing DNA extraction methods. The future goal of this project will be to develop a platform that will accept a sample and automate all steps of sample lysis and ultra-long DNA extraction without user interaction. The proposed system would simplify workflow, lower cost and reduce variability.
