Development of a novel DNA detection system for real-time detection of DNA hybridization

• Published in: Current applied physics Vol. 6; no. 4; pp. 669 - 674
• Main Authors: Kim, Do-Kyun, Tamiya, Eiichi, Kwon, Young-Soo
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.07.2006
• Subjects: DNA chip microarray; DNA hybridization; Evanescent field microscopy; DNA chip microarray; DNA hybridization; Evanescent field microscopy; 87.15.Rn
• ISSN: 1567-1739; 1878-1675
• DOI: 10.1016/j.cap.2005.04.017

With the recent development in DNA hybridization technology, it is now possible to directly analyze oligonucleotide hybridization events with a variety of DNA detection systems in a rapid and simple way. Hybridization of the immobilized oligonucleotides with target oligonucleotides in solution will lead to changes in the mass, electrical charge, or optical properties of the immobilized oligonucleotide layer, which can be detected by microgravimetric, potentiometric, amperometric or optical transducers. In this study, we propose a new DNA detection system of the probe oligonucleotide hybridization based on evanescent field method. First, we have fabricated a DNA chip microarray. The 5′-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray. The particles with immobilized oligonucleotides were arranged by random fluidic self-assembly on the pattern chips through hydrophobic interaction. Second, we have detected DNA hybridization using evanescent field microscopy. The hybridization with fluorescence conjugated target oligonucleotides was elicited by interaction with the evanescent field microscopy. With the evanescent field excitation and real-time detection method described here, we suggest that a very sharp discrimination of bulk fluorescence against surface excitation in combination with high excitation intensities can be achieved. Moreover, monitoring DNA hybridization in a large analytic concentration range and various mismatching conditions were also performed in a rapid and simple procedure.