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Nucleic Acids Research, 2002, Vol. 30, No. 12 e61
© 2002 Oxford University Press

Label-free hybridization detection of a single nucleotide mismatch by immobilization of molecular beacons on an agarose film

Hong Wang, Jiong Li, Heping Liu, Quanjun Liu, Qian Mei, Yijin Wang, Jijun Zhu, Nongyue He and Zuhong Lu*

Chien-Shiung Wu Laboratory, Department of Biomedical Engineering, Southeast University, Nanjing, China

We developed a new technique to immobilize a set of molecular beacons on an agarose film-coated slide and found that it has the ability to identify a single nucleotide difference in label-free DNA targets. The annealing properties, specificity and hybridization dynamics of the present technique were compared with those of the conventional technique that directly immobilizes molecular beacons on a planar glass slide. It is demonstrated that the molecular beacon array on an agarose film has high quench efficiency, an excellent discrimination ratio for single nucleotide mismatches and a short detection time. We hypothesize that such a low fluorescence background and high specificity molecular beacon array will find practical applications in label-free, high-throughput mutation analysis and disease diagnosis.

* To whom correspondence should be addressed. Tel: +86 25 3619983; Fax: +86 25 7712719; Email: zhlu{at}seu.edu.cn


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