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The use of DRAQ5 to monitor intracellular DNA in Escherichia coli by flow cytometry

2010-07Journal article Open access
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dc.contributor.authorSilva, Filomena
dc.contributor.authorLourenço, Olga
dc.contributor.authorVaz, Cidália Pina
dc.contributor.authorRodrigues, Acácio Gonçalves
dc.contributor.authorQueiroz, João
dc.contributor.authorDomingues, F.C.
dc.date.accessioned2019-12-19T16:12:50Z
dc.date.available2019-12-19T16:12:50Z
dc.date.issued2010-07
dc.description.abstractFlow cytometry provides a rapid and high-content multiparameter analysis of individual microorganisms within a population. In the past years, several fluorescent stains were developed in order to monitor DNA content distribution and cell-cycle phases, mainly in eukaryotic cells. Recently, due to its low detection limits, several of these fluorescent stains were also applied to prokaryotic cells. In this study, the ability of a novel far-red fluorescent stain DRAQ5 in assessing intracellular DNA content distribution in Escherichia coli DH5alpha was evaluated. The results showed that a DRAQ5-labelled live E. coli suspension can be obtained by incubation of 1 x 10(6) cells/mL with 5 microM DRAQ5 in PBS buffer supplemented with EDTA (pH = 7.4) during 30 min at 37 degrees C. Flow cytometric analysis of fixed E. coli cells revealed that ethanol should be used in detriment of glutaraldehyde for DRAQ5 labelling. After the analysis of RNase and DNase digested samples, DRAQ5 was proven to be a specific DNA labelling stain. The present study demonstrates that the use of DRAQ5 as a DNA-labelling stain provides an easy assessment of intracellular DNA content and cell-cycle phases in gram-negative bacteria such as E. coli.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1007/s10895-010-0636-ypt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/7996
dc.language.isoengpt_PT
dc.subjectFlow cytometrypt_PT
dc.subjectEscherichia colipt_PT
dc.subjectDNA contentpt_PT
dc.subjectDRAQ5pt_PT
dc.titleThe use of DRAQ5 to monitor intracellular DNA in Escherichia coli by flow cytometrypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage914pt_PT
oaire.citation.issue4pt_PT
oaire.citation.startPage907pt_PT
oaire.citation.titleJournal of Fluorescencept_PT
oaire.citation.volume20pt_PT
person.familyNameSilva
person.familyNameLourenço
person.familyNamePina-Vaz
person.familyNameGonçalves Rodrigues
person.familyNameQueiroz
person.familyNameDomingues
person.givenNameFilomena
person.givenNameOlga Maria Marques
person.givenNameCidália
person.givenNameAcácio
person.givenNameJoão
person.givenNameFernanda
person.identifier1396147
person.identifier.ciencia-idF91E-2365-F208
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person.identifier.ciencia-id6B1F-2A7A-A967
person.identifier.ciencia-idC918-6F2C-6E6A
person.identifier.ciencia-id931E-B66D-E341
person.identifier.ciencia-id6514-0498-3E8F
person.identifier.orcid0000-0003-2931-4583
person.identifier.orcid0000-0002-8401-5976
person.identifier.orcid0000-0003-4936-1301
person.identifier.orcid0000-0003-4246-5212
person.identifier.orcid0000-0002-3096-8325
person.identifier.orcid0000-0002-9540-0853
person.identifier.ridA-3091-2013
person.identifier.ridK-1835-2013
person.identifier.ridK-1467-2013
person.identifier.ridL-3104-2014
person.identifier.ridN-7295-2013
person.identifier.scopus-author-id35723100200
person.identifier.scopus-author-id6701760475
person.identifier.scopus-author-id35582337100
person.identifier.scopus-author-id7003705645
person.identifier.scopus-author-id10139481400
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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relation.isAuthorOfPublication.latestForDiscovery21d53653-d069-4f6b-bc02-266a9bdfe62c

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