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Alizarin Red S for Online Pyrophosphate Detection Identified by a Rapid Screening Method

  • We identified Alizarin Red S and other well known fluorescent dyes useful for the online detection of pyrophosphate in enzymatic assays, including the loop mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assays. An iterative screening was used for a selected set of compounds to first secure enzyme compatibility, evaluate inorganic pyrophosphate sensitivity in the presence of manganese as quencher and optimize conditions for an online detection. Of the selected dyes, the inexpensive alizarin red S was found to selectively detect pyrophosphate under LAMP and PCR conditions and is superior with respect to its defined red-shifted spectrum, long shelf life and low toxicity. In addition, the newly identified properties may also be useful in other enzymatic assays which do not generate nucleic acids but are based on inorganic pyrophosphate. Finally, we propose that our screening method may provide a blueprint for rapid screening of compounds for detecting inorganic pyrophosphate.

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Metadaten
Author:Jens Fischbach, Qiuting Loh, Frank Fabian Bier, Theam Soon Lim, Marcus FrohmeORCiDGND, Jörn GlöklerORCiD
URN:urn:nbn:de:kobv:526-opus4-9248
DOI:https://doi.org/10.1038/srep45085
ISSN:2045-2322
Parent Title (English):Scientific Reports
Document Type:Article
Language:English
Year of Publication:2017
Publishing Institution:Technische Hochschule Wildau
Release Date:2017/03/27
Tag:biochemical assay; fluorescent dye; screening
Volume:7
Source:Fischbach, J. et al. Alizarin Red S for Online Pyrophosphate Detection Identified by a Rapid Screening Method. Sci. Rep.7, 45085; doi: 10.1038/srep45085 (2017).
Faculties an central facilities:Fachbereich Ingenieur- und Naturwissenschaften
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 572 Biochemie
Licence (German):Creative Commons - CC BY - Namensnennung 4.0 International
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