AUTOMATED DNA SYNTHESIZER USING TILTED PLATE TECHNOLOGY

本提案的主题是设计，构建和评估一种新型仪器的原型，用于自动化，平行固相合成大阵列寡核苷酸（一批768至12288种化合物），这将对药物基因分型，基因测序和其他寡核苷酸发现过程非常有益。该仪器的主要优点，使其与目前可用的合成器不同，是一种通过离心从试剂和洗涤溶液中分离固体载体的新技术。在该仪器中，寡核苷酸将在离心机的微量滴度板孔中合成。板向离心中心略微倾斜，从而在离心过程中在每个孔中形成口袋，在口袋中收集固体支撑，而液体则从孔中排出。与目前使用的技术相比，这一过程在机械上更简单、更有效，从而能够制造出更坚固、更灵活、更便宜的仪器。产生的寡核苷酸阵列将用于实现自组装阵列的Illumina技术。建议的商业应用：基于倾斜板技术的半自动化或全自动操作模式的合成器将对简化和加速用于遗传和生物医学研究的大型DNA阵列的平行固相合成产生巨大影响。该合成技术将使Illumina的技术有望占据DNA研究和基因分型市场的很大一部分。这个市场目前估计为每年1亿美元。

The subject of this proposal is the design, construction and evaluation of a prototype of a new instrument for the automated, parallel solid-phase synthesis of large arrays of oligonucleotides (768 to 12,288 compounds in one batch), which will be highly beneficial for the drug genotyping, gene sequencing, and other oligonucleotide involving discovery processes. The principal advantage of this instrument, which sets it apart from currently available synthesizers, is a novel technology for the separation of the solid support from reagent and wash solutions by centrifugation. In this instrument, oligonucleotides will be synthesized in the wells of microtiter plates in a centrifuge. The plates are slightly tilted towards the center of centrifugation, thus forming pockets in each well during centrifugation, in which the solid support is collected, while the liquid is expelled from the wells. This process is mechanically simpler and more efficient compared to currently used technologies, thus enabling the manufacture of more robust, flexible, yet less expensive instruments. Produced arrays of oligonucleotides will be utilized for enabling the Illumina technology of self assembled arrays. PROPOSED COMMERCIAL APPLICATIONS: Synthesizers based on the tilted plate technology in semi- or fully automated mode of operation will have an enormous impact on the simplification and acceleration of the parallel solid-phase synthesis of large DNA arrays for genetic and biomedical research. The synthetic technology will enable Illumina's technology, which is expected to capture significant portion of the DNA research and genotyping market. This market is presently estimated at $100 million per annum.