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Int. J. Mol. Sci. 2014, 15(8), 14332-14347; doi:10.3390/ijms150814332

In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against Atrazine

1
Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, PO Box 9530, Morgantown, WV 26506, USA
2
Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, WV 26506, USA
Current address: Memorial Sloan Kettering Cancer Center, Molecular Pharmacology and Chemistry, 1275 York Avenue, Box 425, New York, NY 10065, USA.
*
Author to whom correspondence should be addressed.
Received: 5 June 2014 / Revised: 11 July 2014 / Accepted: 8 August 2014 / Published: 18 August 2014
(This article belongs to the Section Molecular Recognition)
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Abstract

Widespread use of the chlorotriazine herbicide, atrazine, has led to serious environmental and human health consequences. Current methods of detecting atrazine contamination are neither rapid nor cost-effective. In this work, atrazine-specific single-stranded DNA (ssDNA) molecular recognition elements (MRE) were isolated. We utilized a stringent Systematic Evolution of Ligands by Exponential Enrichment (SELEX) methodology that placed the greatest emphasis on what the MRE should not bind to. After twelve rounds of SELEX, an atrazine-specific MRE with high affinity was obtained. The equilibrium dissociation constant (Kd) of the ssDNA sequence is 0.62 ± 0.21 nM. It also has significant selectivity for atrazine over atrazine metabolites and other pesticides found in environmentally similar locations and concentrations. Furthermore, we have detected environmentally relevant atrazine concentrations in river water using this MRE. The strong affinity and selectivity of the selected atrazine-specific ssDNA validated the stringent SELEX methodology and identified a MRE that will be useful for rapid atrazine detection in environmental samples. View Full-Text
Keywords: SELEX; in vitro selection; aptamer; molecular recognition element (MRE); atrazine SELEX; in vitro selection; aptamer; molecular recognition element (MRE); atrazine
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Williams, R.M.; Crihfield, C.L.; Gattu, S.; Holland, L.A.; Sooter, L.J. In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against Atrazine. Int. J. Mol. Sci. 2014, 15, 14332-14347.

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Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
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