An integrated device for monitoring time-dependent in vitro expression from single genes in picolitre droplets

被引：148

作者：

Courtois, Fabienne ^{[1
,2
]}

Olguin, Luis F. ^{[1
,2
]}

Whyte, Graeme ^{[3
]}

Bratton, Daniel ^{[3
]}

Huck, Wilhelm T. S. ^{[3
]}

Abell, Chris ^{[1
]}

Hollfelder, Florian ^{[2
]}

机构：

[1] Univ Cambridge, Dept Chem, Cambridge CB2 1EW, England

[2] Univ Cambridge, Dept Biochem, Cambridge CB2 1GA, England

[3] Univ Cambridge, Dept Chem, Melville Lab Polymer Synth, Cambridge CB2 1EW, England

来源：

CHEMBIOCHEM | 2008年 / 9卷 / 03期

基金：

英国工程与自然科学研究理事会;

关键词：

high-throughput screening; in vitro protein expression; microfluidic devices; microreactors; monoclonal droplets;

D O I：

10.1002/cbic.200700536

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：

Microdroplets have great potential for high-throughput biochemical screening. We report the design of an integrated microfluidic device for droplet formation, incubation and screening. Picolitre water-in-oil droplets can be stored in a reservoir that contains 106 droplets. In this reservoir droplets are stable for at least 6 h, which gives an extended timescale for biochemical experiments. We demonstrate the utility of the system by following the in vitro expression of green fluorescent protein. The high efficiency allows protein expression from a single molecule of DNA template, creating "monoclonal droplets" in which genotype and phenotype are combined in one emulsion compartment.

引用

页码：439 / 446

页数：8

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