Quantitative Microplate Assay for Real-Time Nuclease Kinetics

Utilizing the phenomenon of nucleases exposing oligonucleotide phosphate backbones to phosphatases we present a novel quantitative method for kinetics of nuclease catalysis. Inorganic phosphate released from nuclease products by phosphatases could be quantified in real-time by a fluorescent sensor o...

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Detalles Bibliográficos
Autores principales: Eriksson, Jonas, Langel, Ülo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4839650/
https://www.ncbi.nlm.nih.gov/pubmed/27101307
http://dx.doi.org/10.1371/journal.pone.0154099
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author Eriksson, Jonas
Langel, Ülo
author_facet Eriksson, Jonas
Langel, Ülo
author_sort Eriksson, Jonas
collection PubMed
description Utilizing the phenomenon of nucleases exposing oligonucleotide phosphate backbones to phosphatases we present a novel quantitative method for kinetics of nuclease catalysis. Inorganic phosphate released from nuclease products by phosphatases could be quantified in real-time by a fluorescent sensor of inorganic phosphate. Two different nucleases were employed, showing the versatility of this assay for multiple turnover label-free nuclease studies.
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spelling pubmed-48396502016-04-29 Quantitative Microplate Assay for Real-Time Nuclease Kinetics Eriksson, Jonas Langel, Ülo PLoS One Research Article Utilizing the phenomenon of nucleases exposing oligonucleotide phosphate backbones to phosphatases we present a novel quantitative method for kinetics of nuclease catalysis. Inorganic phosphate released from nuclease products by phosphatases could be quantified in real-time by a fluorescent sensor of inorganic phosphate. Two different nucleases were employed, showing the versatility of this assay for multiple turnover label-free nuclease studies. Public Library of Science 2016-04-21 /pmc/articles/PMC4839650/ /pubmed/27101307 http://dx.doi.org/10.1371/journal.pone.0154099 Text en © 2016 Eriksson, Langel http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Eriksson, Jonas
Langel, Ülo
Quantitative Microplate Assay for Real-Time Nuclease Kinetics
title Quantitative Microplate Assay for Real-Time Nuclease Kinetics
title_full Quantitative Microplate Assay for Real-Time Nuclease Kinetics
title_fullStr Quantitative Microplate Assay for Real-Time Nuclease Kinetics
title_full_unstemmed Quantitative Microplate Assay for Real-Time Nuclease Kinetics
title_short Quantitative Microplate Assay for Real-Time Nuclease Kinetics
title_sort quantitative microplate assay for real-time nuclease kinetics
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4839650/
https://www.ncbi.nlm.nih.gov/pubmed/27101307
http://dx.doi.org/10.1371/journal.pone.0154099
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