Immobilization of DNA on Magnetic Microparticles for Mercury Enrichment and Detection with Flow Cytometry
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Date
2011-04-26
Authors
Huang, Po-Jung Jimmy
Liu, Juewen
Journal Title
Journal ISSN
Volume Title
Publisher
Wiley
Abstract
Mercury detection in water has attracted a lot of research interest due to its highly toxic nature and adverse environmental impact. In particular, the recent discovery of specific binding of HgII to thymine-rich (T-rich) DNA resulting in T-HgII-T base pairs has led to the development of a number of sensors with different signaling mechanisms. However, the majority of such sensors were non-immobilized. Immobilization, on the other hand, allows active mercury adsorption, signal amplification, and sensor regeneration. In this work, we immobilized a thymine-rich DNA on a magnetic microparticle (MMP) surface through biotin–streptavidin interactions. In the presence of HgII, the DNA changes from a random coil structure into a hairpin, upon which SYBR Green I binds to emit green fluorescence. Detection was carried out by using flow cytometry where the fluorescence intensity increased ≈9-fold in the presence of mercury and the binding of mercury reached equilibrium in less than 2 min. The sensor showed a unique sample-volume-dependent fluorescence signal change where a higher fluorescence was obtained with a larger sample volume, suggesting that the particles can actively adsorb HgII. Detection limits of 5 nM (1 ppb) and 14 nM (2.8 ppb) were achieved in pure buffer and in mercury-spiked Lake Ontario water samples, respectively.
Description
This is the peer reviewed version of the following article: Huang, P.-J. J., & Liu, J. (2011). Immobilization of DNA on Magnetic Microparticles for Mercury Enrichment and Detection with Flow Cytometry. Chemistry - A European Journal, 17(18), 5004–5010, which has been published in final form at https://doi.org/10.1002/chem.201002934. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Keywords
DNA, Mercury, Flow cytometry, Immobilization, Fluorescence, Magnetic microparticles