Characterization of an inexpensive, nontoxic, and highly sensitive microarray substrate.

Martin Dufva; Sarunas Petronis; Louise Bjerremann Jensen; Claudia Krag; Claus B V Christensen

Characterization of an inexpensive, nontoxic, and highly sensitive microarray substrate.

Martin Dufva, Sarunas Petronis, Louise Bjerremann Jensen, Claudia Krag, Claus B V Christensen

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

28 Citationer (Scopus)

Abstract

Udgivelsesdato: 2004-Aug

Originalsprog	Engelsk
Tidsskrift	BioTechniques
Vol/bind	37
Udgave nummer	2
Sider (fra-til)	286-92, 294, 296
ISSN	0736-6205
Status	Udgivet - 2004
Udgivet eksternt	Ja

Bibliografisk note

Keywords: Adsorption; Coated Materials, Biocompatible; DNA Probes; Equipment Design; Equipment Failure Analysis; Hardness; Materials Testing; Oligonucleotide Array Sequence Analysis; Porosity; Reproducibility of Results; Sensitivity and Specificity; Sepharose; Surface Properties

Citationsformater

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title = "Characterization of an inexpensive, nontoxic, and highly sensitive microarray substrate.",

abstract = "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 0.1x 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.",

author = "Martin Dufva and Sarunas Petronis and Jensen, {Louise Bjerremann} and Claudia Krag and Christensen, {Claus B V}",

note = "Keywords: Adsorption; Coated Materials, Biocompatible; DNA Probes; Equipment Design; Equipment Failure Analysis; Hardness; Materials Testing; Oligonucleotide Array Sequence Analysis; Porosity; Reproducibility of Results; Sensitivity and Specificity; Sepharose; Surface Properties",

year = "2004",

language = "English",

volume = "37",

pages = "286--92, 294, 296",

journal = "BioTechniques",

issn = "0736-6205",

publisher = "Informa Healthcare",

number = "2",

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TY - JOUR

T1 - Characterization of an inexpensive, nontoxic, and highly sensitive microarray substrate.

AU - Dufva, Martin

AU - Petronis, Sarunas

AU - Jensen, Louise Bjerremann

AU - Krag, Claudia

AU - Christensen, Claus B V

N1 - Keywords: Adsorption; Coated Materials, Biocompatible; DNA Probes; Equipment Design; Equipment Failure Analysis; Hardness; Materials Testing; Oligonucleotide Array Sequence Analysis; Porosity; Reproducibility of Results; Sensitivity and Specificity; Sepharose; Surface Properties

PY - 2004

Y1 - 2004

N2 - 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 0.1x 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.

AB - 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 0.1x 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.

M3 - Journal article

C2 - 15335222

VL - 37

SP - 286-92, 294, 296

JO - BioTechniques

JF - BioTechniques

SN - 0736-6205

IS - 2

ER -