Parallel DNA Assembling on MicroChip

描述（由申请人提供）：DNA作为分子工具的使用越来越多，已经将DNA的从头合成技术带回了现代生物学的中心舞台。我们建议开发一种强大的超高通量（UHT）技术，能够以当前技术的约1%的成本和时间进行高保真合成DNA的基因组规模生产。我们的第一阶段计划将证明所提出的方法的可行性，该方法包括使用组装脱氧寡核苷酸（oligo）的退火和连接在捕获微芯片上平行合成长DNA序列阵列。我们的第一阶段具体目标是：1：开发一种在芯片上并行合成长DNA序列阵列的方法; 2：开发对提高DNA合成的效率和保真度至关重要的监测和纠错方法。与现有的DNA合成方法相比，所提出的固相平行合成技术具有两个显著的优点：（i）它具有允许同时组装数千个或更多个DNA序列的潜力;（ii）它是可以监测和优化单个步骤和单个合成的过程。如果成功的话，人们可以期待在长DNA序列制备方面迈出重要的一步，类似于将寡核苷酸和肽合成从溶液转移到固相的进步。一个可靠的、小型化的、强大的过程将取代目前缓慢的、昂贵的、仍然很大程度上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.