A microfluidic oligonucleotide synthesizer

被引:68
作者
Lee, Cheng-Chung [1 ,3 ]
Snyder, Thomas M. [1 ,2 ]
Quake, Stephen R. [1 ,2 ]
机构
[1] Stanford Univ, Howard Hughes Med Inst, Stanford, CA 94305 USA
[2] Stanford Univ, Dept Bioengn, Stanford, CA 94305 USA
[3] CALTECH, Dept Bioengn, Pasadena, CA 91125 USA
基金
美国国家科学基金会;
关键词
GENE SYNTHESIS; DEOXYNUCLEOSIDE PHOSPHORAMIDITES; NUCLEOTIDE CHEMISTRY; DEVICE; DEOXYOLIGONUCLEOTIDES; DETRITYLATION;
D O I
10.1093/nar/gkq092
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 similar to 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 similar to 200 bp in length.
引用
收藏
页码:2514 / 2521
页数:8
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