Electroactive intercalators for DNA analysis on microchip electrophoresis.
Miniaturized analytical systems, especially microchip CE (MCE), are becoming a promising tool for analytical purposes including DNA analysis. These microdevices require a sensitive and miniaturizable detection system such as electrochemical detection (ED). Several electroactive DNA intercalators, including the organic dye methylene blue (MB), anthraquinone derivatives, and the metal complexes Fe(phen)3 2+ and Ru(phen)3 2+, have been tested for using in combination with thermoplastic olefin polymer of amorphous structure (Topas) CE-microchips and ED. Two end-channel approaches for integration of gold wire electrodes in CE-ED microchip were used. A 250 microm diameter gold wire was manually aligned at the outlet of the separation channel. A new approach based on a guide channel for integration of 100 and 50 microm diameter gold wire has been also developed in order to reduce the background current and the baseline noise level. Modification of gold wire electrodes has been also tested to improve the detector performance. Application of MCE-ED for ssDNA detection has been studied and demonstrated for the first time using the electroactive dye MB. Electrostatic interaction between cationic MB and anionic ssDNA was used for monitoring the DNA on microchips. Thus, reproducible calibration curves for ssDNA were obtained. This study advances the feasibility of direct DNA analysis using CE-microchip with ED.
Castaño-Alvarez
Mario
M
Departamento de Química Física y Analítica, Universidad de Oviedo, Asturias, Spain.
Fernández-Abedul
M Teresa
MT
Costa-García
Agustín
A
D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000032:analysis*; D046209:Electrophoresis, Microchip / Q000379:methods*; D007364:Intercalating Agents / Q000737:chemistry*
D004277:DNA, Single-Stranded; D007364:Intercalating Agents