Automated Large and Mid Scale Nucleic Acid Preparation

DESCRIPTION (provided by applicant): This research involves the development a disposable purification cassette and bench-top electrophoretic processing instrument that will purify one milligram or more of plasmid DNA from recombinant culture in 60 minutes.The purification method to be employed in this work is a derivative of proven electrophoretic separation technology that our company developed for automated mini-prep DNA purification. Data obtained prior to Phase I demonstrated that up to 0.3 milligramsof highly pure plasmid DNA could be automatically purified from 50 mls of liquid bacterial culture loaded directly into a prototype cassette. This preliminary data indicates that cassettes could be made to process up to 150 mls of culture and thus yield 1mg or more of plasmid DNA. Phase I work will establish the proof-of-concept for the product. Aims of Phase I include: optimization of the cassette design to determine the limitations of the scaled-up process, and optimization of the electrophoretic voltage program to reduce the run time to one hour, without yield loss, and with minimal user steps. The process will also be tested for purification of genomic DNA from bacterial and mammalian cells. A microprocessor control circuit will be constructed in Phase I that will be the basis of the actual processing instrument to be developed in Phase II. The products from this research will be faster and far easier to use than any instrument or manual kit currently available. The disposables of this product are projected to cost 10 per sample, while the bench-top instrument will have a price of ~ 2000. The method has essentially no moving parts. This work will greatly reduce the labor and cost required to purify milligram amounts of plasmid DNA. Automated purification of plasmid and genomic DNA participates in a multi- hundred million-dollar nucleic acid purification market. The product has the potential to generate 10-20 million per year in revenue. PUBLIC HEALTH RELEVANCE: The work will create a bench-top instrument and disposable cassettes that will allow for fully automated purification of milligram quantities of plasmid and genomic DNA. This technology will be significantly less expensive and far less time consumptive that any existing instrument or manual kit available.