Department of Bioengineering, California Institute of Technology, Pasadena, CA 91125, USA.
Nucleic Acids Res. 2010 May;38(8):2514-21. doi: 10.1093/nar/gkq092. Epub 2010 Feb 21.
De novo gene and genome synthesis enables the design of any sequence without the requirement of a pre-existing template as in traditional genetic engineering methods. The ability to mass produce synthetic genes holds great potential for biological research, but widespread availability of de novo DNA constructs is currently hampered by their high cost. In this work, we describe a microfluidic platform for parallel solid phase synthesis of oligonucleotides that can greatly reduce the cost of gene synthesis by reducing reagent consumption (by 100-fold) while maintaining a approximately 100 pmol synthesis scale so there is no need for amplification before assembly. Sixteen oligonucleotides were synthesized in parallel on this platform and then successfully used in a ligation-mediated assembly method to generate DNA constructs approximately 200 bp in length.
从头基因和基因组合成使设计任何序列成为可能,而无需像传统遗传工程方法那样依赖于预先存在的模板。大规模生产合成基因的能力为生物研究带来了巨大的潜力,但从头 DNA 构建体的广泛可用性目前受到其高成本的阻碍。在这项工作中,我们描述了一种用于寡核苷酸平行固相合成的微流控平台,该平台可以通过减少试剂消耗(降低 100 倍)来大大降低基因合成的成本,同时保持大约 100pmol 的合成规模,因此在组装之前无需进行扩增。在该平台上平行合成了 16 个寡核苷酸,然后成功地用于连接介导的组装方法,以生成大约 200bp 长的 DNA 构建体。