In this work, we provided the evidence of an alternative DNA detection methodology, in the DNA microarray domain, based on a Universal Microarray approach. Despite the classic hybridization of an oligonucleotide immobilized onto a proper surface with a DNA target extracted from a biological sample and amplified by a PCR reaction, here we reported an alternative method based on the usage of generic oligonucleotides microarray sequences, coupled to special probes contained in the hybridization buffer, at end user level. These special probes present one region complementary to the oligonucleotide on the surface (microarray) and the other part recognizing a certain DNA target sequence. These special probes act like a bridge connection and the final hybridization leads to a three-units complex formation. We demonstrated that the hybridization occurs, and therefore the DNA target recognition, although with less efficiency, that is reasonable because of the thermodynamics constrains in the formation of the unstable three-units complex. However, signal detection is still comparable with standard hybridization.

A bridge-like solution for Universal Microarray applications

Alessi E.;Petralia S.
Ultimo
2016-01-01

Abstract

In this work, we provided the evidence of an alternative DNA detection methodology, in the DNA microarray domain, based on a Universal Microarray approach. Despite the classic hybridization of an oligonucleotide immobilized onto a proper surface with a DNA target extracted from a biological sample and amplified by a PCR reaction, here we reported an alternative method based on the usage of generic oligonucleotides microarray sequences, coupled to special probes contained in the hybridization buffer, at end user level. These special probes present one region complementary to the oligonucleotide on the surface (microarray) and the other part recognizing a certain DNA target sequence. These special probes act like a bridge connection and the final hybridization leads to a three-units complex formation. We demonstrated that the hybridization occurs, and therefore the DNA target recognition, although with less efficiency, that is reasonable because of the thermodynamics constrains in the formation of the unstable three-units complex. However, signal detection is still comparable with standard hybridization.
Fluorescent detection
Lab-on-Chip
Nucleic acids
Universal Microarray
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.11769/519623
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