標題: Neutralized chimeric DNA probe for detection of single nucleotide polymorphism on surface plasmon resonance biosensor
作者: Huang, Chun-Jen
Lin, Zhong-En
Yang, Yuh-Shyong
Chan, Hardy Wai-Hong
Chen, Wen-Yih
生物科技學系
Department of Biological Science and Technology
關鍵字: SNP genotyping;SPR imaging;Label-free detection;Ethylated DNA;DNA analogue
公開日期: 15-Jan-2018
摘要: An implementation of neutralized chimeric DNA oligomer as a probe for sensitive detection of single nucleotide polymorphisms (SNPs) on a surface plasmon resonance imaging sensor is investigated. The chimeric DNA oligomer was synthesized in a conventional DNA synthesizer, containing neutral nucleotides with a methylated phosphate group. The secondary structures and melting points of the chimeric DNA fragment and its complexes with perfect-matched and single-mismatched complementary DNA molecules were examined by using circular dichroism and UV vis spectroscopy in comparison with the native probe DNA counterpart. The results indicate that the chimeric DNA complexes can form a B-form structure and exhibit high thermostability. Moreover, the hybridization and discrimination efficiency of the chimeric probe DNA for the SNP genotyping were verified by using the SPRi sensor under different experimental conditions. The data reveal the effects of the ionic strength and operation temperature on the selectivity of the chimeric probe DNA for the SNP detection. The hybridization condition with a low ionic strength and high temperature allows the chimeric probe DNA distinguishing perfect-matched and single-mismatched target DNA molecules to the best extent, likely due to the reduced electrostatic repulsive force and presence of the additional methyl group on the backbone. Consequently, the direct and label-free detection with the SPR technique and neutralized chimeric probe DNA can be realized for the SNP genotyping by optimizing the operation condition and sequence design.
URI: http://dx.doi.org/10.1016/j.bios.2017.07.052
http://hdl.handle.net/11536/143948
ISSN: 0956-5663
DOI: 10.1016/j.bios.2017.07.052
期刊: BIOSENSORS & BIOELECTRONICS
Volume: 99
起始頁: 170
結束頁: 175
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