Sorted libraries of error-free long oligonucleotides

描述：合成生物学充满了希望，从发现和生产新药，靶向治疗修改生物体如酵母或大肠杆菌或创造全新的，更高的作物产量，二氧化碳封存到生物能源项目。然而，这些目标很难达到，主要是由于De Novo基因合成和从化学合成DNA到合成染色体的新控制和调节代谢途径的合成困难。就像任何可逆的化学反应一样，通过依次添加单体（A，C，G，T）来合成DNA片段，每一步的产率都低于100%（典型的逐步产率为~97-99%），因此要合成的DNA长度越长，最终的纯产物的比例越小。误差的消除

DESCRIPTION: Synthetic Biology is full of promises ranging from discovery and production of new drugs, targeted therapies modifying organisms such as yeast or E. coli or creating totally new ones, higher crop yields, CO2 sequestration to bio-energy projects. However, these goals have been difficult to reach primarily due to the difficulty of De Novo gene synthesis and synthesis of new control and regulatory metabolic pathways all the way to synthetic chromosomes from chemically synthesized DNA. Just like any reversible chemical reaction DNA fragment synthesis by sequentially adding the monomers (A,C,G,T) gives less than 100% yield at each step( typical stepwise yields are ~97-99%) as a result the longer the length of DNA to be synthesized the smaller is the fraction of pure product at the end. Elimination of the errors take significant time and money. The goal of this project is to provide error free long oligonucleotides (100 to 300 mers) at a cost lower than the impure DNA fragments of today to unlock the potential of synthetic biology. We are proposing to make libraries of clonally amplified and sequence verified long oligonucleotides. Our long term objectives are to implement a commercial service of affordable custom synthesis of sequence-verified long oligonucleotide libraries. During Phase I, we will 1) to demonstrate that single oligonucleotide molecules can be clonally amplified and sequence verified, 2) to demonstrate that beads bearing clonal amplifications can be captured on a microarray bearing sequence-specific probes and 3) to demonstrate that sequence verified oligonucleotides enable gene assembly with ten fold reduced error rate. During Phase II, we will 1) to develop a protocol/device to normalize the number of each oligonucleotide surveyed from a library, 2) to establish a standard operating procedure for the production of large libraries of sequence-verified oligonucleotides and 3) to determine the maximum oligonucleotide length that could be offered as a commercial product.