基于聚合酶链反应的重叠寡核苷酸-寡聚物不对称延伸基因合成在寡核苷酸延伸模拟器的支持下获得了1 kbp的dsDNA。

IF 2.2 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS
Yasunori Nishida, Kotetsu Kayama, Taichi Endoh, Kiwamu Hanazono, Gerry Amor Camer, Daiji Endoh
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引用次数: 0

摘要

在寡核苷酸延伸(AESOE)模拟器的支持下,通过不对称延伸,采用“寡聚物单向连接法”合成了1 kbp的DNA片段。本研究对10个黄病毒基因组的41组不同基因组片段和31个细菌16s rRNA片段进行了试验,这些片段的大小从500个碱基到1.0 kbp不等。合成基因的产生在所有这些组合中都是成功的。该合成方法有三步:第一步是七链AESOE,第二步是连接第一步的400碱基片段,第三步是最终扩增。我们目前的方法具有很高的可重复性,并且可能不再需要优化低聚物设计。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA.

We formulated a method to synthesize 1 kbp DNA fragments using 'oligomer unidirectional joining method' via asymmetric extension supported by a simulator for oligonucleotide extension (AESOE). In this study, trials were conducted on 41 sets of different genomic pieces of ten flaviviral genomes, and 31 bacterial 16s rRNA fragments with sizes ranging from 500 bases to 1.0 kbp. Synthetic gene production was found to be successful in all those sets. The synthesis method has three steps: the first step is a seven-linked AESOE, the second step is the linking of the 400-base fragments from the first step, and the third step is the final amplification. Our present approach is highly reproducible and may no longer require optimization of oligomer design.

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来源期刊
BioTechniques
BioTechniques 工程技术-生化研究方法
CiteScore
4.40
自引率
0.00%
发文量
68
审稿时长
3.3 months
期刊介绍: BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.
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