Parallel gene synthesis in a microfluidic device.
Parallel gene synthesis in a microfluidic device.
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DOI：
10.1093/nar/gkm121
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发表时间：
2007
影响因子：
14.9
通讯作者：
Jacobson JM
中科院分区：
文献类型：
--
作者：
Kong DS;Carr PA;Chen L;Zhang S;Jacobson JM
由于合成基因和更长的DNA结构在传统分子生物学和新兴的合成生物学领域都有许多强大的应用，所以快速、准确和廉价地合成定制从头DNA结构的能力是研究人员非常希望的。然而，目前从头合成的成本--主要是由试剂和处理成本驱动--是这种技术广泛应用的一个重大障碍。在这项工作中，我们展示了，据我们所知，第一次在微流控环境中进行基因合成。微流控技术的使用大大减少了反应体积以及相应的试剂和处理成本。此外，微流控技术使大量复杂的反应能够并行进行。在这里，我们报告了一种多腔微流控装置的制造及其在几种DNA结构的合成中的应用。在4个500 NL反应器中，以微小的起始寡核苷酸浓度(10-25 NM)平行合成长达1 kb的基因。这样的体积比传统的基因合成低一到两个数量级。通过测序验证了所有目的基因的同源性，结果错误率确定为每560个碱基中有1个。
The ability to synthesize custom de novo DNA constructs rapidly, accurately and inexpensively is highly desired by researchers, as synthetic genes and longer DNA constructs are enabling to numerous powerful applications in both traditional molecular biology and the emerging field of synthetic biology. However, the current cost of de novo synthesis—driven largely by reagent and handling costs—is a significant barrier to the widespread availability of such technology. In this work, we demonstrate, to our knowledge, the first gene synthesis in a microfluidic environment. The use of microfluidic technology greatly reduces reaction volumes and the corresponding reagent and handling costs. Additionally, microfluidic technology enables large numbers of complex reactions to be performed in parallel. Here, we report the fabrication of a multi-chamber microfluidic device and its use in carrying out the syntheses of several DNA constructs. Genes up to 1 kb in length were synthesized in parallel at minute starting oligonucleotide concentrations (10–25 nM) in four 500 nl reactors. Such volumes are one to two orders of magnitude lower than those utilized in conventional gene synthesis. The identity of all target genes was verified by sequencing, and the resultant error rate was determined to be 1 per 560 bases.
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影响因子：
56.9
作者：
CARUTHERS, MH
通讯作者：
CARUTHERS, MH
影响因子：
56.9
作者：
Unger, MA;Chou, HP;Quake, SR
通讯作者：
Quake, SR
影响因子：
7.4
作者：
Liu, J;Hansen, C;Quake, SR
通讯作者：
Quake, SR
影响因子：
14.9
作者：
Hoover, DM;Lubkowski, J
通讯作者：
Lubkowski, J
DOI：
10.1101/sqb.1986.051.01.032
发表时间：
1986-01-01
期刊：
COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY
影响因子：
--
作者：
MULLIS, K;FALOONA, F;ERLICH, H
通讯作者：
ERLICH, H