Parallel DNA Assembling on MicroChip

描述(申请人提供)：DNA作为分子工具的使用越来越多，使DNA从头合成技术重新回到现代生物学的中心阶段。我们建议开发一种强大的超高通量(UHT)技术，能够以当前技术约1%的成本和时间生产高保真合成DNA的基因组规模。我们的第一阶段计划将证明所提出的方法的可行性，该方法包括在捕获芯片上并行合成一组长DNA序列，使用退火法和连接组装脱氧寡核苷酸(寡核苷酸)。我们第一阶段的具体目标是：1：开发一种在芯片上并行合成长DNA序列的方法；2：开发对提高DNA合成的效率和保真度至关重要的监控和纠错方法。与现有的DNA合成方法相比，提出的固相平行合成技术具有两个显著的优点：(I)它有可能同时组装数千或更多的DNA序列；(Ii)它是一个可以监控和优化单个步骤和单个合成的过程。如果成功，人们可以期待在长DNA序列制备方面向前迈出重要的一步，类似于将寡核苷酸和肽合成从溶液转移到固相的进展。然后，一个可靠、小型化、健壮的过程将取代目前缓慢、昂贵且在很大程度上仍不可预测的基因合成过程。

DESCRIPTION (provided by applicant): The increasing use of DNA as a molecular tool has ushered the technology of de novo synthesis of DNA back to the center stage of modern biology. We propose the development of a robust, ultra high-throughput (UHT) technology capable of genome-scale production of high fidelity synthetic DNA at about one percent of the cost and time of the current technologies. Our phase I plan will demonstrate the feasibility of the proposed method that includes parallel synthesis of an array of long DNA sequences on a capture microchip using annealing and ligation of assembling deoxyoligonucleotides (oligos). Our Phase I Specific Aims are: 1: Developing a method for parallel synthesis of an array of long DNA sequences on chip; 2: Developing monitoring and error correction methods essential for improving the efficiency and fidelity of the DNA synthesis. The proposed solid phase parallel synthesis technology has two significant advantages over the existing methods of DNA synthesis: (i) it has the potential to allow simultaneous assembling of thousands or more DNA sequences; (ii) it is a process that individual steps and individual synthesis can be monitored and optimized. If successful, one can expect a significant step forward in long DNA sequence preparation, similar to the advancement by moving oligonucleotide and peptide synthesis from solution to solid phase. A reliable, miniaturized, robust process will then replace the current slow, expensive, and still largely unpredictable gene synthesis processes.