Development of a graphene oxide-based assay for the sequence-specific detection of double-stranded DNA molecules.
Graphene oxide (GO) is a promising material for the development of cost-effective detection systems. In this work, we have devised a simple and rapid GO-based method for the sequence-specific identification of DNA molecules generated by PCR amplification. The csp genes of Escherichia coli, which sha...
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2017-01-01
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doaj-540fe5c4eefb4e07b2e66013d44ff1b12020-11-25T01:49:56ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01128e018395210.1371/journal.pone.0183952Development of a graphene oxide-based assay for the sequence-specific detection of double-stranded DNA molecules.Anna Maria GiuliodoriAnna BrandiShivaram KotlaFrancesco PerrozziRoberto GunnellaLuca OttavianoRoberto SpurioAttilio FabbrettiGraphene oxide (GO) is a promising material for the development of cost-effective detection systems. In this work, we have devised a simple and rapid GO-based method for the sequence-specific identification of DNA molecules generated by PCR amplification. The csp genes of Escherichia coli, which share a high degree of sequence identity, were selected as paradigm DNA templates. All tested csp genes were amplified with unlabelled primers, which can be rapidly removed at the end of the PCR taking advantage of the preferential binding to GO of single-stranded versus duplex DNA molecules. The amplified DNAs (targets) were heat-denatured and hybridized to a fluorescently-labelled single strand oligonucleotide (probe), which recognizes a region of the target DNAs displaying sequence variability. This interaction is extremely specific, taking place with high efficiency only when target and probe show perfect or near perfect matching. Upon GO addition, the unbound fraction of the probe was captured and its fluorescence quenched by the GO's molecular properties. On the other hand, the probe-target complexes remained in solution and emitted a fluorescent signal whose intensity was related to their degree of complementarity.http://europepmc.org/articles/PMC5574608?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anna Maria Giuliodori Anna Brandi Shivaram Kotla Francesco Perrozzi Roberto Gunnella Luca Ottaviano Roberto Spurio Attilio Fabbretti |
spellingShingle |
Anna Maria Giuliodori Anna Brandi Shivaram Kotla Francesco Perrozzi Roberto Gunnella Luca Ottaviano Roberto Spurio Attilio Fabbretti Development of a graphene oxide-based assay for the sequence-specific detection of double-stranded DNA molecules. PLoS ONE |
author_facet |
Anna Maria Giuliodori Anna Brandi Shivaram Kotla Francesco Perrozzi Roberto Gunnella Luca Ottaviano Roberto Spurio Attilio Fabbretti |
author_sort |
Anna Maria Giuliodori |
title |
Development of a graphene oxide-based assay for the sequence-specific detection of double-stranded DNA molecules. |
title_short |
Development of a graphene oxide-based assay for the sequence-specific detection of double-stranded DNA molecules. |
title_full |
Development of a graphene oxide-based assay for the sequence-specific detection of double-stranded DNA molecules. |
title_fullStr |
Development of a graphene oxide-based assay for the sequence-specific detection of double-stranded DNA molecules. |
title_full_unstemmed |
Development of a graphene oxide-based assay for the sequence-specific detection of double-stranded DNA molecules. |
title_sort |
development of a graphene oxide-based assay for the sequence-specific detection of double-stranded dna molecules. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2017-01-01 |
description |
Graphene oxide (GO) is a promising material for the development of cost-effective detection systems. In this work, we have devised a simple and rapid GO-based method for the sequence-specific identification of DNA molecules generated by PCR amplification. The csp genes of Escherichia coli, which share a high degree of sequence identity, were selected as paradigm DNA templates. All tested csp genes were amplified with unlabelled primers, which can be rapidly removed at the end of the PCR taking advantage of the preferential binding to GO of single-stranded versus duplex DNA molecules. The amplified DNAs (targets) were heat-denatured and hybridized to a fluorescently-labelled single strand oligonucleotide (probe), which recognizes a region of the target DNAs displaying sequence variability. This interaction is extremely specific, taking place with high efficiency only when target and probe show perfect or near perfect matching. Upon GO addition, the unbound fraction of the probe was captured and its fluorescence quenched by the GO's molecular properties. On the other hand, the probe-target complexes remained in solution and emitted a fluorescent signal whose intensity was related to their degree of complementarity. |
url |
http://europepmc.org/articles/PMC5574608?pdf=render |
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