In Phase II, we will develop, and test a simple, cost-effective device that allows bacterial growth in any type of 96 or 384 well microtiter plates or deep well blocks without the use of expensive equipment. The product simplifies bacterial growth in tightly spaced microtiter plates, yet still result in robust culture with DNA yields greater than equivalent volume grown in Erlenmeyer or larger shaker test tubes. In phase II, we will further develop and optimize an aeration/agitation prototype device that was successfully experimented with in Phase I. This prototype uses a controlled stream of pro-heated, sterile, humidified air that is passed through the culture medium to simultaneously accomplish aeration, heating and agitation of the tightly packed cultures. The prototype works well with small and large culture volumes ranging from 50 mu l to 1.5 ml, and thus could be used for growth in 96 or 384 deep well plates. The DNA yield from culture volumes as low as 300 mu l was sufficient quantity to perform several automated DNA sequencing reactions resulting in long, high accuracy reads. The proposed product will be inexpensive to construct and will operate in a completely self-contained, compact manner without an external incubator, shaker, stirrer or oxygen source as are used with other devices that accomplish this task. We believe, based on prototypes developed in Phase I, the device can be sold for less than $2000. This price compares favorably to competitor products costing $7500 to $30,000. The product will greatly enhance the ability to grow and prepare DNA from recombinant cultures, thereby improving the quality and throughput of large-scale DNA sequencing and analysis efforts. It will also be highly useful in molecular biology labs that are increasingly faced with growth and preparation of larger numbers of recombinant DNA samples. The product address the molecular biology researchers found in over 40,000 labs throughout the world. It has a market potential of $200-300 million dollars and will save significant research dollars both in private companies and grant-funded non-profit labs.
