Characterization of an inexpensive, nontoxic, and highly sensitive microarray substrate
| Autor: | Martin Dufva, Claudia Krag, Claus Christensen, Sarunas Petronis, Louise Bjerremann Jensen |
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| Předmět: |
Analyte
Materials science Microscope Surface Properties Scanning electron microscope Sensitivity and Specificity General Biochemistry Genetics and Molecular Biology law.invention chemistry.chemical_compound Coated Materials Biocompatible Hardness law Materials Testing Sodium dodecyl sulfate Oligonucleotide Array Sequence Analysis Chromatography Sepharose Hybridization probe Reproducibility of Results Substrate (chemistry) Equipment Design Molecular biology Equipment Failure Analysis chemistry Agarose Adsorption DNA Probes Porosity Layer (electronics) Biotechnology |
| Zdroj: | ResearcherID Scopus-Elsevier |
| Popis: | An agarose film has been proposed as an efficient substrate for producing microarrays. The original film preparation procedure was simplified significantly by grafting the agarose layer directly onto unmodified microscope glass slides instead of aminated glass slides, and the blocking procedure was replaced with a wash in 0.1× standard saline citrate (SSC) and 0.5% sodium dodecyl sulfate (SDS) without decreasing the performance of the produced microarrays. Characterization of the grafted agarose film using atomic force microscopy (AFM) and scanning electron microscopy (SEM) showed that the agarose film had a 10-fold increase in surface roughness compared to glass and that the interior of the agarose film was porous, with pore sizes between 100–500 nm. A comparison of hybridization on aldehyde-activated agarose-coated microarray slides and commercial amino-reactive microarray slides showed that aldehyde-activated agarose-coated slides had the highest signal-to-noise ratio of 850, suggesting that the aldehyde-activated agarose microarray slides are suitable in applications where analytes have a wide concentration range. By immobilizing the DNA probes using ultraviolet (UV) light, the signal-to-noise ratio was further increased to 3000 on the agarose microarray slides. The specificity of the UV cross-linked DNA probes was demonstrated using 21 and 25 bp long capture probes, enabling discrimination of target molecules differing in only one base. |
| Databáze: | OpenAIRE |
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