Oligonucleotide chip assay for quantification of gamma ray-induced single strand breaks.
Identifieur interne : 002779 ( Main/Exploration ); précédent : 002778; suivant : 002780Oligonucleotide chip assay for quantification of gamma ray-induced single strand breaks.
Auteurs : Hyeon A. Ki [Corée du Sud] ; Min Jung Kim ; Sukdeb Pal ; Joon Myong SongSource :
- Journal of pharmaceutical and biomedical analysis [ 0731-7085 ] ; 2009.
Descripteurs français
- KwdFr :
- ADN simple brin (analyse), ADN simple brin (effets des radiations), ADN simple brin (génétique), Altération de l'ADN (effets des radiations), Biotine (métabolisme), Biotinylation, Calibrage, Colorants fluorescents (métabolisme), Facteurs temps, Fluorescéine (métabolisme), Oligonucléotides (génétique), Propriétés de surface, Rayons gamma, Relation dose-effet des radiations, Silicium (), Séquençage par oligonucléotides en batterie ().
- MESH :
- analyse : ADN simple brin.
- effets des radiations : ADN simple brin, Altération de l'ADN.
- génétique : ADN simple brin, Oligonucléotides.
- métabolisme : Biotine, Colorants fluorescents, Fluorescéine.
- Biotinylation, Calibrage, Facteurs temps, Propriétés de surface, Rayons gamma, Relation dose-effet des radiations, Silicium, Séquençage par oligonucléotides en batterie.
English descriptors
- KwdEn :
- Biotin (metabolism), Biotinylation, Calibration, DNA Damage (radiation effects), DNA, Single-Stranded (analysis), DNA, Single-Stranded (genetics), DNA, Single-Stranded (radiation effects), Dose-Response Relationship, Radiation, Fluorescein (metabolism), Fluorescent Dyes (metabolism), Gamma Rays, Oligonucleotide Array Sequence Analysis (methods), Oligonucleotides (genetics), Silicon (chemistry), Surface Properties, Time Factors.
- MESH :
- chemical , analysis : DNA, Single-Stranded.
- chemical , chemistry : Silicon.
- chemical , genetics : DNA, Single-Stranded, Oligonucleotides.
- chemical , metabolism : Biotin, Fluorescein, Fluorescent Dyes.
- methods : Oligonucleotide Array Sequence Analysis.
- radiation effects : DNA Damage, DNA, Single-Stranded.
- Biotinylation, Calibration, Dose-Response Relationship, Radiation, Gamma Rays, Surface Properties, Time Factors.
Abstract
An oligonucleotide chip assay was designed for direct quantification of single strand breaks (SSBs) induced by gamma-ray irradiation. The oligonucleotides used were 20-mers, which were short enough to produce only a single strand break within a single oligonucleotide. The two ends of the oligonucleotides were labeled with fluorescein and biotin, respectively. The biotinylated ends of the oligonucleotides were immobilized on a silicon wafer chip treated with (3-aminopropyl)triethoxysilane (APTES), glutaraldehyde, and avidin. The DNA fragments cleaved by gamma-ray irradiation were detected by a laser-induced fluorescence (LIF) detection system. The gamma-ray-induced SSBs were quantified using a calibration curve (fluorescence intensity versus gamma-ray dose) without the need for complicated mathematical calculation based on gel-based separation. The experimentally determined gamma-ray-induced SSBs yield was almost equal to the theoretical value derived from gel electrophoresis of plasmid DNAs and DNA surface coverage.
DOI: 10.1016/j.jpba.2008.11.031
PubMed: 19128912
Affiliations:
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Le document en format XML
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<author><name sortKey="Kim, Min Jung" sort="Kim, Min Jung" uniqKey="Kim M" first="Min Jung" last="Kim">Min Jung Kim</name>
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<series><title level="j">Journal of pharmaceutical and biomedical analysis</title>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Biotin (metabolism)</term>
<term>Biotinylation</term>
<term>Calibration</term>
<term>DNA Damage (radiation effects)</term>
<term>DNA, Single-Stranded (analysis)</term>
<term>DNA, Single-Stranded (genetics)</term>
<term>DNA, Single-Stranded (radiation effects)</term>
<term>Dose-Response Relationship, Radiation</term>
<term>Fluorescein (metabolism)</term>
<term>Fluorescent Dyes (metabolism)</term>
<term>Gamma Rays</term>
<term>Oligonucleotide Array Sequence Analysis (methods)</term>
<term>Oligonucleotides (genetics)</term>
<term>Silicon (chemistry)</term>
<term>Surface Properties</term>
<term>Time Factors</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ADN simple brin (analyse)</term>
<term>ADN simple brin (effets des radiations)</term>
<term>ADN simple brin (génétique)</term>
<term>Altération de l'ADN (effets des radiations)</term>
<term>Biotine (métabolisme)</term>
<term>Biotinylation</term>
<term>Calibrage</term>
<term>Colorants fluorescents (métabolisme)</term>
<term>Facteurs temps</term>
<term>Fluorescéine (métabolisme)</term>
<term>Oligonucléotides (génétique)</term>
<term>Propriétés de surface</term>
<term>Rayons gamma</term>
<term>Relation dose-effet des radiations</term>
<term>Silicium ()</term>
<term>Séquençage par oligonucléotides en batterie ()</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>DNA, Single-Stranded</term>
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<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Silicon</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Single-Stranded</term>
<term>Oligonucleotides</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Biotin</term>
<term>Fluorescein</term>
<term>Fluorescent Dyes</term>
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<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ADN simple brin</term>
</keywords>
<keywords scheme="MESH" qualifier="effets des radiations" xml:lang="fr"><term>ADN simple brin</term>
<term>Altération de l'ADN</term>
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<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN simple brin</term>
<term>Oligonucléotides</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Oligonucleotide Array Sequence Analysis</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Biotine</term>
<term>Colorants fluorescents</term>
<term>Fluorescéine</term>
</keywords>
<keywords scheme="MESH" qualifier="radiation effects" xml:lang="en"><term>DNA Damage</term>
<term>DNA, Single-Stranded</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Biotinylation</term>
<term>Calibration</term>
<term>Dose-Response Relationship, Radiation</term>
<term>Gamma Rays</term>
<term>Surface Properties</term>
<term>Time Factors</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Biotinylation</term>
<term>Calibrage</term>
<term>Facteurs temps</term>
<term>Propriétés de surface</term>
<term>Rayons gamma</term>
<term>Relation dose-effet des radiations</term>
<term>Silicium</term>
<term>Séquençage par oligonucléotides en batterie</term>
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<front><div type="abstract" xml:lang="en">An oligonucleotide chip assay was designed for direct quantification of single strand breaks (SSBs) induced by gamma-ray irradiation. The oligonucleotides used were 20-mers, which were short enough to produce only a single strand break within a single oligonucleotide. The two ends of the oligonucleotides were labeled with fluorescein and biotin, respectively. The biotinylated ends of the oligonucleotides were immobilized on a silicon wafer chip treated with (3-aminopropyl)triethoxysilane (APTES), glutaraldehyde, and avidin. The DNA fragments cleaved by gamma-ray irradiation were detected by a laser-induced fluorescence (LIF) detection system. The gamma-ray-induced SSBs were quantified using a calibration curve (fluorescence intensity versus gamma-ray dose) without the need for complicated mathematical calculation based on gel-based separation. The experimentally determined gamma-ray-induced SSBs yield was almost equal to the theoretical value derived from gel electrophoresis of plasmid DNAs and DNA surface coverage.</div>
</front>
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<affiliations><list><country><li>Corée du Sud</li>
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<name sortKey="Song, Joon Myong" sort="Song, Joon Myong" uniqKey="Song J" first="Joon Myong" last="Song">Joon Myong Song</name>
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<country name="Corée du Sud"><region name="Région capitale de Séoul"><name sortKey="Ki, Hyeon A" sort="Ki, Hyeon A" uniqKey="Ki H" first="Hyeon A" last="Ki">Hyeon A. Ki</name>
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