Nanobind Hierarchical Silica Lamella for High Molecular Weight DNA Extraction

DESCRIPTION provided by applicant As new genetic analysis methods are developed rising emphasis is being placed on high quality sample preparation and the ability to obtain large quantities of high molecular weight and high purity DNA In DNA sequencing rd generation technologies offer long read lengths up to kb and require high quality isolated DNA In gene therapy and DNA vaccines high binding capacity could be extremely useful in isolating industrial quantities of long plasmids In genetic analysis of extremely rare sequences or functional analysis of large chromosomal rearrangements high molecular weight input DNA reduces the chance that fragmentation will occur in a region of interest In Phase I we created a novel thermoplastic silica substrate containing a hierarchical layering of microscale folds and nanoscale silica lamella called Nanobind Through the combined effects of minimized shear force exposure and high surface area we are able to extract vast amounts X more of high molecular weight genomic DNA andgt kb using a simple bind wash and elute protocol that is both fast andlt min and inexpensive $ per substrate In this Phase II SBIR we will develop Nanobind technologies to address areas where critical limitations exist with current extraction methods First we will create a facile method for high MW extraction by developing a magnetic Nanobind substrate Second current automated extraction systems rely exclusively on beads and columns and tend to fragment DNA to andlt kb We will develop MagNanobind HTP a well format for high throughput high MW DNA extraction that is compatible with a variety of existing automated extraction instruments Third we will develop a microvolume Nanobind cartridge to perform small volume extractions using clinical samples andlt L or andlt ng Finally we will validate the Nanobind DNA RNA extraction platform by comparing against microparticle spin column and precipitation methods using common genetic and epigenetic assays Nanobind could immediately impact the DNA extraction market as it uniquely combines the speed and ease of microparticles and spin columns with performance of phenol chloroform PUBLIC HEALTH RELEVANCE As new genetic analysis methods are developed rising emphasis is being placed on high quality sample preparation This project will develop a new silica nanomaterial for nucleic acid isolation that is fast and easy yet results in large quantitie of high quality DNA and RNA Such a method could greatly reduce the cost of sample preparation while improving the quality of data from genetic research