The Human Genome Project requires the rapid synthesis of very large numbers of oligonucleotide primers for polymerase chain reaction amplification of human DNA, and for use as probes for sequence-tagged sites. This proposal relates to the invention of a system for the rapid automatic multiple- parallel synthesis of oligonucleotides. In our system, a series of sequential reactions are carried out in parallel, with a proper and pre- programmed nucleotide added to each reaction train, with reagent streams accurately apportioned and separated without the use of separate pumps, with individual reagents effectively segmented, with positive methods for moving small volumes through the system and for displacing one reagent quantitatively with the next, and with reagent volumes kept to a minimum. In addition, the final cleaved product will be purified chromatographically on-line and delivered in a dry form. Two versions of the system are proposed, one using a conventional particulate solid-phase support, the second based on new technology which allows the solid support to be a porous film. With the present design 50 or more primers can be made in parallel. The system will be more compact and cheaper than present synthesizers, and can be built on either a micro- or macro-scale.
