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Nucleic acid and protein extraction from electropermeabilized E. coli cells on a microfluidic chip

2013-09-26Journal article Restricted access
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dc.contributor.authorMatos, T.
dc.contributor.authorSenkbeil, S.
dc.contributor.authorMendonça, António
dc.contributor.authorQueiroz, João
dc.contributor.authorKutter, Jörg
dc.contributor.authorBulow, L.
dc.date.accessioned2022-08-24T08:40:31Z
dc.date.available2022-08-24T08:40:31Z
dc.date.issued2013-09-26
dc.description.abstractDue to the extensive use of nucleic acid and protein analysis of bacterial samples, there is a need for simple and rapid extraction protocols for both plasmid DNA and RNA molecules as well as reporter proteins like the green fluorescent protein (GFP). In this report, an electropermeability technique has been developed which is based on exposing E. coli cells to low voltages to allow extraction of nucleic acids and proteins. The flow-through electropermeability chip used consists of a microfluidic channel with integrated gold electrodes that promote cell envelope channel formation at low applied voltages. This will allow small biomolecules with diameters less than 30 A to rapidly diffuse from the permeabilized cells to the surrounding solution. By controlling the applied voltage, partial and transient to complete cell opening can be obtained. By using DC voltages below 0.5 V, cell lysis can be avoided and the transiently formed pores can be closed again and the cells survive. This method has been used to extract RNA and GFP molecules under conditions of electropermeability. Plasmid DNA could be recovered when the applied voltage was increased to 2 V, thus causing complete cell lysis.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationMatos, T., Senkbeil, S., Mendonça, A., Queiroz, J. A., Kutter, J. P., & Bulow, L. (2013). Nucleic acid and protein extraction from electropermeabilized E. coli cells on a microfluidic chip. Analyst, 138(24), 7347-7353. doi:10.1039/c3an01576apt_PT
dc.identifier.doi10.1039/c3an01576apt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/12309
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherRSC Publishingpt_PT
dc.relationDEVELOPMENT AND ANALYSIS OF NOVEL CHROMATOGRAPHIC MATERIALS USEFUL FOR PROTEINS AND PEPTIDES CHARACTERIZATION
dc.relation.publisherversionhttps://pubs.rsc.org/en/content/articlelanding/2013/an/c3an01576apt_PT
dc.subjectNucleic acidpt_PT
dc.subjectE. coli cellspt_PT
dc.subjectProtein - Electropermeabilizedpt_PT
dc.titleNucleic acid and protein extraction from electropermeabilized E. coli cells on a microfluidic chippt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleDEVELOPMENT AND ANALYSIS OF NOVEL CHROMATOGRAPHIC MATERIALS USEFUL FOR PROTEINS AND PEPTIDES CHARACTERIZATION
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/FARH/SFRH%2FBD%2F47934%2F2008/PT
oaire.citation.endPage7353pt_PT
oaire.citation.issue24pt_PT
oaire.citation.startPage7347pt_PT
oaire.citation.titleThe Analystpt_PT
oaire.citation.volume138pt_PT
oaire.fundingStreamFARH
person.familyNameMendonça
person.familyNameQueiroz
person.familyNameKutter
person.givenNameAntónio
person.givenNameJoão
person.givenNameJörg
person.identifier.ciencia-id2419-6E20-F958
person.identifier.ciencia-id931E-B66D-E341
person.identifier.orcid0000-0001-8069-0440
person.identifier.orcid0000-0002-3096-8325
person.identifier.orcid0000-0003-1065-1985
person.identifier.ridI-3093-2012
person.identifier.ridL-3104-2014
person.identifier.ridE-3256-2010
person.identifier.scopus-author-id56890866600
person.identifier.scopus-author-id7003705645
person.identifier.scopus-author-id8871525100
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.embargofctCopyright cedido à editora no momento da publicação: The Royal Society of Chemistry 2013pt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
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relation.isAuthorOfPublication.latestForDiscoveryc798c033-c160-4f99-a8ad-04416923943f
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