Pyrosequencing on a glass surface

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dc.contributor.authorAlmeida, Ana V.cs
dc.contributor.authorManz, Andreascs
dc.contributor.authorNeužil, Pavelcs
dc.coverage.issue6cs
dc.coverage.volume16cs
dc.date.accessioned2021-12-08T15:54:39Z
dc.date.available2021-12-08T15:54:39Z
dc.date.issued2016-02-16cs
dc.description.abstractWe demonstrate the use of open-surface microuidics to sequence DNA by pyrosequencing at the plain hydrophobically coated surface of a microscope glass cover slip. This method offers signicant advantages in terms of instrument size, simplicity, disposability, and functional integration, particularly when combined with the broad and exible capabilities of open-surface microuidics. The DNA was incubated on superparamagnetic particles and placed on a hydrophobically coated glass substrate. The particles with bound DNA were moved using magnetic force through microliter-sized droplets covered with mineral oil to prevent water evaporation from the droplets. These droplets served as reaction “stations” performing pyrosequencing as well as washing stations. The resequencing protocol with 34-mer single-stranded DNA (ssDNA) was used to determine the reaction performance. The de novo sequencing was performed with 51-mer and 81-mer ssDNA. The method can be integrated with previously shown sample preparation and PCR into a single sample-to-answer system on a plain glass surface.en
dc.formattextcs
dc.format.extent1063-1071cs
dc.format.mimetypeapplication/pdfcs
dc.identifier.citationLAB ON A CHIP. 2016, vol. 16, issue 6, p. 1063-1071.en
dc.identifier.doi10.1039/c6lc00114acs
dc.identifier.issn1473-0197cs
dc.identifier.other122201cs
dc.identifier.urihttp://hdl.handle.net/11012/203127
dc.language.isoencs
dc.publisherRoyal Society of Chemistrycs
dc.relation.ispartofLAB ON A CHIPcs
dc.relation.urihttps://pubs.rsc.org/en/content/articlelanding/2016/LC/C6LC00114Acs
dc.rightsCreative Commons Attribution 3.0 Unportedcs
dc.rights.accessopenAccesscs
dc.rights.sherpahttp://www.sherpa.ac.uk/romeo/issn/1473-0197/cs
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/cs
dc.subjectDNA sequence determinationen
dc.subjectpyrosequencingen
dc.subjectdropletsen
dc.subjectmagnetic particlesen
dc.subjectopen-surface microfluidicsen
dc.subjectvirtual reaction chambersen
dc.titlePyrosequencing on a glass surfaceen
dc.type.driverarticleen
dc.type.statusPeer-revieweden
dc.type.versionpublishedVersionen
sync.item.dbidVAV-122201en
sync.item.dbtypeVAVen
sync.item.insts2021.12.08 16:54:39en
sync.item.modts2021.12.08 16:14:34en
thesis.grantorVysoké učení technické v Brně. Středoevropský technologický institut VUT. Chytré nanonástrojecs
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