The development of improved techniques for DNA sequencing is necessary to achieve the time and cost goals of the Human Genome Project. One important component of assembling a sequencing reaction is to step along a DNA fragment using an oligonucleotide to prime the next part of the sequencing reaction (primer walking). This type of oligonucleotide priming only requires about 5 picomoles of oligomer; however, conventional oligonucleotide synthesizers produce more than 10,000 times that amount. The proposed project involves the development of a synthesizer that can produce smaller oligonucleotide amounts appropriate for sequencing at costs that are more than an order of magnitude lower than conventional systems. In addition to making primer walking more cost effective, the almost immediate access (few hours) to a wide array of very inexpensive oligonucleotides in a research laboratory would make feasible new experimental approaches to DNA sequencing, PCR analysis and diagnostics, mutagenesis and gene synthesis. The proposed project combines recent advancements in low cost parallel synthesis of DNA oligonucleotides with advanced computer-controlled machinery design in an effort to produce a cost effective method of synthesizing a large number of oligonucleotides. The Phase II project will involve the design, construction and testing of several versions of a prototype instrument.
