A double-emulsion microfluidic platform for in vitro green fluorescent protein expression
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Wu, Nan; Oakeshott, John Graham; Easton, Christopher; Peat, Thomas S; Surjadi, Regina; Zhu, Yonggang
Description
Microfluidic droplet technology has gained popularity due to the advantages over conventional emulsion techniques and capabilities for a wide range of applications. In this paper, the development of a simple microfluidic-based double-emulsion system is reported. Such a system could be potentially used for in vitro protein synthesis. The system involves a two-step process to make water-in-oil-in-water (W/O/W) emulsions. A PMMA microchip is used for the formation of water-in-oil (W/O)...[Show more]
dc.contributor.author | Wu, Nan | |
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dc.contributor.author | Oakeshott, John Graham | |
dc.contributor.author | Easton, Christopher | |
dc.contributor.author | Peat, Thomas S | |
dc.contributor.author | Surjadi, Regina | |
dc.contributor.author | Zhu, Yonggang | |
dc.date.accessioned | 2015-12-10T22:51:08Z | |
dc.identifier.issn | 0960-1317 | |
dc.identifier.uri | http://hdl.handle.net/1885/58911 | |
dc.description.abstract | Microfluidic droplet technology has gained popularity due to the advantages over conventional emulsion techniques and capabilities for a wide range of applications. In this paper, the development of a simple microfluidic-based double-emulsion system is reported. Such a system could be potentially used for in vitro protein synthesis. The system involves a two-step process to make water-in-oil-in-water (W/O/W) emulsions. A PMMA microchip is used for the formation of water-in-oil (W/O) single-emulsion droplets. Then, the single-emulsion droplets are transported to a PDMS/glass microchip to make the W/O/W double-emulsion droplets. The system was first characterized by detecting fluorescein sodium salt as a model dye in the internal aqueous droplets using laser-induced fluorescence. The effect of the flow rates of the internal aqueous phase and outer continuous aqueous phase on the formation of the double-emulsion droplets is investigated to provide information for system optimization. On-chip storage of double-emulsion droplets is also investigated to allow for protein synthesis from a PCR-generated DNA template using either commercial in vitro transcription and translation kits or crude Escherichia coli S30 extracts. In vitro expression of the green fluorescent protein is successfully demonstrated in this system. | |
dc.publisher | IOP Publishing | |
dc.source | Journal of Micromechanics and Microengineering | |
dc.subject | Keywords: Aqueous droplets; Aqueous phase; Conventional emulsions; DNA-template; Fluorescein sodiums; Green fluorescent protein; Green fluorescent protein expression; In-vitro; Laser induced fluorescence; Microfluidic platforms; On-chip storage; Protein synthesis; | |
dc.title | A double-emulsion microfluidic platform for in vitro green fluorescent protein expression | |
dc.type | Journal article | |
local.description.notes | Imported from ARIES | |
local.identifier.citationvolume | 21 | |
dc.date.issued | 2011 | |
local.identifier.absfor | 030401 - Biologically Active Molecules | |
local.identifier.ariespublication | u4005981xPUB464 | |
local.type.status | Published Version | |
local.contributor.affiliation | Wu, Nan, College of Physical and Mathematical Sciences, ANU | |
local.contributor.affiliation | Oakeshott, John Graham, CSIRO Division of Entomology | |
local.contributor.affiliation | Easton, Christopher, College of Physical and Mathematical Sciences, ANU | |
local.contributor.affiliation | Peat, Thomas S, CSIRO Materials Science and Engineering | |
local.contributor.affiliation | Surjadi, Regina, CSIRO Materials Science and Engineering | |
local.contributor.affiliation | Zhu, Yonggang, CSIRO Materials Science and Engineering | |
local.description.embargo | 2037-12-31 | |
local.bibliographicCitation.issue | 5 | |
local.bibliographicCitation.startpage | 054032/1 | |
local.bibliographicCitation.lastpage | 7 | |
local.identifier.doi | 10.1088/0960-1317/21/5/054032 | |
local.identifier.absseo | 970103 - Expanding Knowledge in the Chemical Sciences | |
dc.date.updated | 2016-02-24T10:24:15Z | |
local.identifier.scopusID | 2-s2.0-79955846836 | |
local.identifier.thomsonID | 000289986600033 | |
Collections | ANU Research Publications |
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