A single square voltage pulse is used to achieve the selective covalent bonding of DNA probes to a functionalized thin-film surface (SiO2) on a plastic substrate. Significant covalent bonding occurs with only 100 ns of pulse duration, representing a decrease in immobilization time of more than ten orders of magnitude from the 2 h needed without electric field. A systematic study of the effect of the duration and rise/fall times of the voltage pulse on the immobilization of DNA probe molecules is presented. Successful hybridization of the DNA probes immobilized using this technique with a complementary target strand was achieved.
In this work, a procedure to immobilize DNA probes on a microarray patterned on a flexible plastic substrate is developed. The method involves the chemical activation of a thin film surface, the introduction of amine functionality via a silanization step, the coupling of an adequate crosslinker and finally the immobilization of the DNA probe. The response of different thin-film materials and plastic substrates to the immobilization procedure is discussed. The DNA probes immobilized in the patterned pixels were then allowed to hybridize with complementary target DNA labeled with a fluorescent molecule. A prototype array of thin film pixels of SiO2 functionalized by silanization deposited over a polyimide substrate is demonstrated.
Electric fields generated by single square and sinusoidal voltage pulses with amplitudes below 2 V were used to assist the covalent immobilization of single-stranded, thiolated DNA probes, onto a chemically functionalized SiO2 surface and to assist the specific hybridization of single-stranded DNA targets with immobilized complementary probes. The single-stranded immobilized DNA probes were either covalently immobilized (chemisorption) or electrostatically adsorbed (physisorption) to a chemically functionalized surface. Comparing the speed of electric field assisted immobilization and hybridization with the corresponding control reactions (without electric field), an increase of several orders of magnitude is observed, with the reaction timescaled down from 1 to 2 h to a range between 100 ns and 1 ms. The influence of the shape of the voltage pulse (square versus sinusoidal) and its duration were studied for both immobilization and hybridization reactions. The results show that pulsed electric fields are a useful tool to achieve temporal and spatial control of surface immobilization and hybridization reactions of DNA.
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