Quantitative Microplate Assay for Real-Time Nuclease Kinetics

Jonas Eriksson; Ülo Langel

doi:10.1371/journal.pone.0154099

Quantitative Microplate Assay for Real-Time Nuclease Kinetics

PLoS One. 2016 Apr 21;11(4):e0154099. doi: 10.1371/journal.pone.0154099. eCollection 2016.

Authors

Jonas Eriksson¹, Ülo Langel¹

Affiliation

¹ Department of Neurochemistry, Stockholm University, Stockholm, Sweden.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Catalysis
Endonucleases / chemistry*
Kinetics

Substances

Endonucleases

Grants and funding

This work was supported by grants to ÜL from the Swedish Research Council for Natural Sciences (621-2011-5902), the Swedish Research Council for Medical Research (K2012-66X-21148-04-5) and the Swedish Cancer Society (CAN 2012/251). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.