The effects of glass surfaces and probe GC content on signal intensities of a 60-mer diagnostic microarray.
Identifieur interne : 001992 ( PubMed/Checkpoint ); précédent : 001991; suivant : 001993The effects of glass surfaces and probe GC content on signal intensities of a 60-mer diagnostic microarray.
Auteurs : Xiaoyang Mo ; Qinghua Wu [République populaire de Chine] ; Junjian Hu ; Wenli Ma [République populaire de Chine] ; Min Wei [République populaire de Chine] ; Wuzhou Yuan ; Yuequn Wang ; Yongqin Li ; Yun Deng ; Xiushan WuSource :
- Annals of microbiology [ 1590-4261 ] ; 2008.
Abstract
The effects of glass surfaces and probe GC content on signal intensities of a 60-mer diagnostic microarray were studied. Twelve virus-specific oligonucleotide probes for severe acute respiratory syndrome coronavirus (SARS-CoV) were divided into a high GC content group (≥ 50%) and a low GC content group (< 50%), and spotted onto four different chemically-modified glass surfaces: a poly-amine coating activated by 1,4-phenylene diisothiocyanate (Poly-Amine surface), an acrylic acid-co-acrylamide copolymer coating activated by 1-(3-dimethylamino propyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (AACA-Copolymer surface), a commercial Corning CMT-GAPS amino surface, and a Telechem SuperAmine amino surface. RNA samples from cultured SARS-CoV strain were labelled using direct cDNA labelling with restriction display in a single colour format. The background-subtracted signal intensities were analysed using two-way analysis of variance. The effects of glass surfaces on background-subtracted signal intensities were significant (p=0.003). Multiple comparisons showed that differences existed mainly between the AACA-Copolymer surface and the other glass surfaces, and that the AACA-Copolymer surface had the highest background-subtracted signal intensity. The probe GC content had no significant effect on signal intensities in the narrow range of GC content represented (p=0.07). The results suggested that the AACA-Copolymer surface may be a novel choice of microorganism survey based on long oligonucleotide microarray.
DOI: 10.1007/BF03175336
PubMed: 32226355
Affiliations:
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<front><div type="abstract" xml:lang="en">The effects of glass surfaces and probe GC content on signal intensities of a 60-mer diagnostic microarray were studied. Twelve virus-specific oligonucleotide probes for severe acute respiratory syndrome coronavirus (SARS-CoV) were divided into a high GC content group (≥ 50%) and a low GC content group (< 50%), and spotted onto four different chemically-modified glass surfaces: a poly-amine coating activated by 1,4-phenylene diisothiocyanate (Poly-Amine surface), an acrylic acid-co-acrylamide copolymer coating activated by 1-(3-dimethylamino propyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (AACA-Copolymer surface), a commercial Corning CMT-GAPS amino surface, and a Telechem SuperAmine amino surface. RNA samples from cultured SARS-CoV strain were labelled using direct cDNA labelling with restriction display in a single colour format. The background-subtracted signal intensities were analysed using two-way analysis of variance. The effects of glass surfaces on background-subtracted signal intensities were significant (p=0.003). Multiple comparisons showed that differences existed mainly between the AACA-Copolymer surface and the other glass surfaces, and that the AACA-Copolymer surface had the highest background-subtracted signal intensity. The probe GC content had no significant effect on signal intensities in the narrow range of GC content represented (p=0.07). The results suggested that the AACA-Copolymer surface may be a novel choice of microorganism survey based on long oligonucleotide microarray.</div>
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<Abstract><AbstractText>The effects of glass surfaces and probe GC content on signal intensities of a 60-mer diagnostic microarray were studied. Twelve virus-specific oligonucleotide probes for severe acute respiratory syndrome coronavirus (SARS-CoV) were divided into a high GC content group (≥ 50%) and a low GC content group (< 50%), and spotted onto four different chemically-modified glass surfaces: a poly-amine coating activated by 1,4-phenylene diisothiocyanate (Poly-Amine surface), an acrylic acid-co-acrylamide copolymer coating activated by 1-(3-dimethylamino propyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (AACA-Copolymer surface), a commercial Corning CMT-GAPS amino surface, and a Telechem SuperAmine amino surface. RNA samples from cultured SARS-CoV strain were labelled using direct cDNA labelling with restriction display in a single colour format. The background-subtracted signal intensities were analysed using two-way analysis of variance. The effects of glass surfaces on background-subtracted signal intensities were significant (p=0.003). Multiple comparisons showed that differences existed mainly between the AACA-Copolymer surface and the other glass surfaces, and that the AACA-Copolymer surface had the highest background-subtracted signal intensity. The probe GC content had no significant effect on signal intensities in the narrow range of GC content represented (p=0.07). The results suggested that the AACA-Copolymer surface may be a novel choice of microorganism survey based on long oligonucleotide microarray.</AbstractText>
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<name sortKey="Wang, Yuequn" sort="Wang, Yuequn" uniqKey="Wang Y" first="Yuequn" last="Wang">Yuequn Wang</name>
<name sortKey="Wu, Xiushan" sort="Wu, Xiushan" uniqKey="Wu X" first="Xiushan" last="Wu">Xiushan Wu</name>
<name sortKey="Yuan, Wuzhou" sort="Yuan, Wuzhou" uniqKey="Yuan W" first="Wuzhou" last="Yuan">Wuzhou Yuan</name>
</noCountry>
<country name="République populaire de Chine"><noRegion><name sortKey="Wu, Qinghua" sort="Wu, Qinghua" uniqKey="Wu Q" first="Qinghua" last="Wu">Qinghua Wu</name>
</noRegion>
<name sortKey="Ma, Wenli" sort="Ma, Wenli" uniqKey="Ma W" first="Wenli" last="Ma">Wenli Ma</name>
<name sortKey="Wei, Min" sort="Wei, Min" uniqKey="Wei M" first="Min" last="Wei">Min Wei</name>
</country>
</tree>
</affiliations>
</record>
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