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Quantitative Microplate Assay for Real-Time Nuclease Kinetics.

ABSTRACT: 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.

SUBMITTER: Eriksson J

PROVIDER: S-EPMC4839650 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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Quantitative Microplate Assay for Real-Time Nuclease Kinetics.

Eriksson Jonas J Langel Ülo Ü

PloS one 20160421 4

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. ...[more]

PMID: 27101307

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