This work will create innovative products that allow rapid, low cost, automated, genomic DNA purification from several sample types and concentrations. This novel technology uses a disposable cassette device and low cost, reusable processing cradle to automatically purify DNA by self-contained, semi-dry electrophoretic methodology. Cassettes can be constructed to contain multiple lanes that allow simultaneous purification of a few, or up to 96 samples in one run. The method requires no moving parts and can be performed, with some sample types, in less than 10 minutes. The projected cost of cassettes is $0.4 - $0.8 per sample, while the portable processing cradle will cost less than $250 for a 1-12 sample version, or $750 for a 96-sample version. This product, if fully realized, could revolutionize the way that DNA is prepared for clinical or reference laboratory diagnostic procedures. It will reduce costs and save labor and materials. Work performed in Phase I with prototypes of the purification cassette and processing devices showed that DNA prepared from blood, bacteria, saliva or tissue is highly pure and can be used directly in PCR amplification, as well as other molecular biology applications. Phase I work also showed that the method could begin with a wide range of sample concentrations, as the starting sample could be diluted 100-500 fold, and method still yielded active DNA template. The purification method utilizes a variation of technology that our company developed for automated plasmid DNA purification, combined with technology for running agarose gels without running buffer. Phase II work will result in the working prototypes for the cassettes and processing cradle instruments.
Aims of Phase II are to: 1) Further optimize the electrophoretic voltage programs. 2) Improve the lower detection limit of bacterial and mammalian cells, including purification of DNA from single mammalian cells. 3) Improve the lysis chemistry. 4) Determine the scope of activity of the purified DNA. 5) Develop a protocol for small volumes of blood. 6) Develop the 12 and 96-well processing cradle instruments. 7) Finalize the 2, 6, 12 and 24-well cassette design and contract for the construction of injection molds. 8) Draft an instruction manual and product specifications. Our company will be able to manufacture and sell instruments and cassettes developed from this work directly after Phase II. The market potential for these products is estimated to be ~$50 million. The products are also applicable to clinical labs and possibly doctor's office settings.
This proposal will create innovative products that allow rapid, low cost, automated, multi-sample genomic DNA purification from many sample types and concentrations. The novel technology uses disposable, recyclable cassette device and low cost processing cradle to automatically purify DNA by electrophoretic means, without running buffer baths or platinum electrodes.