Detection of deletion mutations in DNA using water-soluble cationic fluorescent thiophene copolymer |
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Authors: | Hongliang Guan |
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Affiliation: | Key Laboratory of Analytical Chemistry for Biology and Medicine, College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, PR China |
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Abstract: | In this contribution, we designed a fluorescent thiophene copolymer to detect insertion/deletion mutation in DNA by doping aldehyde group in the main chain. The fluorescence of the copolymer could be dramatically quenched on the addition of single-stranded DNA (ssDNA) via strong electrostatic interactions and electronic/energy transfer. Although the complementary ssDNA made the fluorescence recover, the hydrogen bonds and chemical coupling also played a significant role between the unpaired bases and aldehyde group, which could differentiate the subtle differences in such mutant DNA. The influence of buffer pH, concentration of NaCl, heating time and the temperature was systemically investigated and the proposed method was then successfully applied to detect real sample. With the respect to the linearity, limit of detection precision, specificity, this procedure could provide sensitive methodologies for the rapid detection and identification of nucleic acids. |
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Keywords: | Conjugated copolymer Fluorescence Chemical coupling Hydrogen bond Lifetime |
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