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.
Author: | Jens Fischbach, Qiuting Loh, Frank Fabian Bier, Theam Soon Lim, Marcus FrohmeORCiDGND, Jörn GlöklerORCiD |
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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 |