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dc.contributor.authorKatzmeier, Florian-
dc.contributor.authorAufinger, Lukas-
dc.contributor.authorDupin, Aurore-
dc.contributor.authorQuintero, Jorge-
dc.contributor.authorLenz, Matthias-
dc.contributor.authorBauer, Ludwig-
dc.contributor.authorKlumpe, Sven-
dc.contributor.authorSherpa, Dawafuti-
dc.contributor.authorDürr, Benedikt-
dc.contributor.authorHonemann, Maximilian-
dc.contributor.authorStyazhkin, Igor-
dc.contributor.authorSimmel, Friedrich C.-
dc.contributor.authorHeymann, Michael-
dc.date.accessioned2020-03-13T09:26:59Z-
dc.date.available2020-03-13T09:26:59Z-
dc.date.issued2019de
dc.identifier.issn1932-6203-
dc.identifier.other169245479X-
dc.identifier.urihttp://nbn-resolving.de/urn:nbn:de:bsz:93-opus-ds-108085de
dc.identifier.urihttp://elib.uni-stuttgart.de/handle/11682/10808-
dc.identifier.urihttp://dx.doi.org/10.18419/opus-10791-
dc.description.abstractPoint-of-care testing (POCT) in low-resource settings requires tools that can operate independently of typical laboratory infrastructure. Due to its favorable signal-to-background ratio, a wide variety of biomedical tests utilize fluorescence as a readout. However, fluorescence techniques often require expensive or complex instrumentation and can be difficult to adapt for POCT. To address this issue, we developed a pocket-sized fluorescence detector costing less than $15 that is easy to manufacture and can operate in low-resource settings. It is built from standard electronic components, including an LED and a light dependent resistor, filter foils and 3D printed parts, and reliably reaches a lower limit of detection (LOD) of ≈ 6.8 nM fluorescein, which is sufficient to follow typical biochemical reactions used in POCT applications. All assays are conducted on filter paper, which allows for a flat detector architecture to improve signal collection. We validate the device by quantifying in vitro RNA transcription and also demonstrate sequence-specific detection of target RNAs with an LOD of 3.7 nM using a Cas13a-based fluorescence assay. Cas13a is an RNA-guided, RNA-targeting CRISPR effector with promiscuous RNase activity upon recognition of its RNA target. Cas13a sensing is highly specific and adaptable and in combination with our detector represents a promising approach for nucleic acid POCT. Furthermore, our open-source device may be used in educational settings, through providing low cost instrumentation for quantitative assays or as a platform to integrate hardware, software and biochemistry concepts in the future.en
dc.language.isoende
dc.relation.uridoi:10.1371/journal.pone.0220091de
dc.rightsinfo:eu-repo/semantics/openAccessde
dc.subject.ddc570de
dc.subject.ddc610de
dc.titleA low-cost fluorescence reader for in vitro transcription and nucleic acid detection with Cas13aen
dc.typearticlede
ubs.fakultaetEnergie-, Verfahrens- und Biotechnikde
ubs.fakultaetFakultätsübergreifend / Sonstige Einrichtungde
ubs.institutInstitut für Biomaterialien und biomolekulare Systemede
ubs.institutFakultätsübergreifend / Sonstige Einrichtungde
ubs.publikation.seiten17de
ubs.publikation.sourcePLOS ONE 14 (2019), No. 12, e0220091de
ubs.publikation.typZeitschriftenartikelde
Enthalten in den Sammlungen:04 Fakultät Energie-, Verfahrens- und Biotechnik

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9 iGem2017.pdfKatzmeier et al. 20192,21 MBAdobe PDFÖffnen/Anzeigen
pone.0220091.s001.pdfKatzmeier et al. 2019 Appendix S1128,81 kBAdobe PDFÖffnen/Anzeigen
pone.0220091.s002.pngKatzmeier et al. 2019 Fig S11,59 MBimage/pngÖffnen/Anzeigen
pone.0220091.s003.tifKatzmeier et al. 2019 Fig S28,47 MBTIFFÖffnen/Anzeigen
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pone.0220091.s009.pdfKatzmeier et al. 2019 File S345,81 kBAdobe PDFÖffnen/Anzeigen


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