High fidelity low cost manufacture of synthetic DNA

DESCRIPTION (provided by applicant): The long term objective of this project is to develop technology to provide low-cost ( 90% and to reliably assemble these into genes. We aim to lower the cost of building block synthesis 3- to 10-fold by combining advanced chemistry with alternative engineering allowing a reduction in reaction volumes and reagent concentrations. It is essential for this application that this reduced cost is accompanied by an increase in quality: lower quality oligonucleotides assemble into genes that have increased error frequencies and thus require additional downstream manipulations and sequence verification, increasing the overall cost of production. We will therefore optimize synthesis on a 2- dimensional support to improve the stepwise coupling efficiency to >99.5%; in Phase I we demonstrated the feasibility of this by obtaining coupling efficiencies of 99.9% using modified chemistries and procedures. Small scale, low cost, high coupling efficiency oligonucleotide synthesis will allow greater flexibility in oligonucleotide gene building block design, remove the need for oligonucleotide purification prior to assembly into complete genes, reduce sequence confirmation costs and shorten order to delivery times. To achieve this, in Phase II we will develop an automated 96 channel instrument for synthesizing oligonucleotides with >90% purity. The machine will be integrated into our existing high throughput gene production line at DNA2.0. A low cost high fidelity oligonucleotide manufacturing process may have additional applications in related areas including diagnostic microarrays, real time PCR and RNAi-reagents.