【摘 要】
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Applications of microchip-synthesized oligonucleotides for gene synthesis are limited by their low yield, low quality, and high complexity.A simple and less
【机 构】
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Departmentoflifescience,UniversityofScienceandTechnologyofChina
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Applications of microchip-synthesized oligonucleotides for gene synthesis are limited by their low yield, low quality, and high complexity.A simple and less equipment-dependent de novo gene synthesis method, integrated with high-throughput error-removal and multiple gene assembly procedure, was developed for large-scale, low-cost, synthesis of high-quality DNA based on microchip-synthesized oligonucleotides.The initialed synthesis oligos pool in a chip was corrected using high-throughput error-removal by immobilized MutS, and then assembled to high-quality fragments.Then the corresponding high-fidelity fragments pool assembly from this error corrected oligos pool was further into multiple full-length gene by MPCA.Using this method, ten gene encoding a lycopene synthesis pathway was synthesis.The error frequency was reduced by 26.3-fold (from 14.47/kb to 0.55/kb).Resulting in a 15.5-fold in the percentage of error-free fragments (from 3.23% to 50%).This method allows the use of the otherwise highly error-prone microchip-synthesized oligonucleotides for de novo gene synthesis without pre-purification.
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