An improved gene synthesis method with asymmetric directions of oligonucleotides designed using a simulation program.
An improved gene synthesis method with asymmetric directions of oligonucleotides designed using a simulation program.
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DOI：
10.2144/btn-2020-0062
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发表时间：
2020-09-01
期刊：
影响因子：
2.7
通讯作者：
Endoh, Daiji
中科院分区：
文献类型：
--
作者：
Kayama, Kotetsu;Hashizume, Hibiki;Endoh, Daiji
基于寡核苷酸扩增的人工基因合成被称为重叠延伸PCR，它产生多种中间合成产物。利用序列低聚物扩展模拟程序，可以调整低聚物的取向和浓度，以减少中间产物的合成，优化DNA合成的全长过程。当低聚物处于“正-反-反-反”方向时，序列低聚物延伸过程的效率最高。具有这种设计方向的低聚物证明了通过重复退火和延伸在最短的时间内（循环次数）产生所需的产物。该方法被命名为非对称延伸，由寡核苷酸延伸模拟器（AESOE）支持，在DNA合成中显示出效率和有效性，具有未来改进和优化应用的潜力。
Artificial gene synthesis based on oligonucleotide augmentationis known as overlap extension PCR which generates a variety of intermediate synthetic products. The orientation and concentration of oligomers can be adjusted to reduce the synthesis of intermediates and optimize the full-length process of DNA synthesis, using a simulation program for serial oligomer extension. The efficiency of the serial oligomer extension process is predicted to be greatest when oligomers are in a 'forward-reverse-reverse-reverse' direction. Oligomers with such designed directions demonstrated generation of the desired product in the shortest time (number of cycles) by repeated annealing and elongation. This method, named Asymmetric Extension supported by a Simulator for Oligonucleotide Extension (AESOE), has shown efficiency and effectiveness withpotentials for future improvements and optimal usage in DNA synthesis.