MersV1, Main, Exploration, bibRecord, 002B58

Guanosine gel for sequence-dependent separation of polymorphic ssDNA.

Identifieur interne : 002B58 ( Main/Exploration ); précédent : 002B57; suivant : 002B59

Guanosine gel for sequence-dependent separation of polymorphic ssDNA.

Auteurs : William S. Case [États-Unis] ; Keren D. Glinert ; Sam Labarge ; Linda B. Mcgown

Source :

Electrophoresis [ 0173-0835 ] ; 2007.

RBID : pubmed:17661319

Descripteurs français

KwdFr :
- ADN simple brin (analyse), Colorants fluorescents, Composition en bases nucléiques, Données de séquences moléculaires, Guanosine monophosphate (), Séquence nucléotidique, Électrochromatographie capillaire.
MESH :
- analyse : ADN simple brin.
- Colorants fluorescents, Composition en bases nucléiques, Données de séquences moléculaires, Guanosine monophosphate, Séquence nucléotidique, Électrochromatographie capillaire.

English descriptors

KwdEn :
- Base Composition, Base Sequence, Capillary Electrochromatography, DNA, Single-Stranded (analysis), Fluorescent Dyes, Guanosine Monophosphate (chemistry), Molecular Sequence Data.
MESH :
- chemical , analysis : DNA, Single-Stranded.
- chemical , chemistry : Guanosine Monophosphate.
- Base Composition, Base Sequence, Capillary Electrochromatography, Fluorescent Dyes, Molecular Sequence Data.

Abstract

The separation of fluorescent-labeled ssDNA fragments of equal length based on differences in sequence was achieved through the use of guanosine gels (G-gels) formed by guanosine-5'-monophosphate (GMP) in capillary gel electrokinetic chromatography (CGEKC) with LIF detection. Baseline resolution was achieved for homodimers and homopentamers of A, T, and C. G-gel CGEKC provided better resolution than CZE, MEKC, or a sieving gel in CGE. Resolution improved with increasing GMP, indicating that the interaction is linked to structural organization of the G-gel. Fluorescence intensity and anisotropy show that the order of interaction with G-gels is T>C>A. We then investigated four conformationally similar, polymorphic 76-mers with A/G substitutions that are utilized in forensic DNA typing. Resolution was achieved by CGEKC but not CZE or CGE. In CGEKC, the negatively charged G-gels and oligonucleotides electromigrate toward the positive inlet while being driven by EOF to the negative outlet. The net forward velocity is the greatest for oligonucleotides most closely associated with the slower, more cumbersome G-gel network. For the 76-mers, resolution increases with increasing difference in guanosine content between strands and, for a given difference, with increasing total guanosines in the strands.

DOI: 10.1002/elps.200700287
PubMed: 17661319

Affiliations:

Le document en format XML

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<affiliation wicri:level="2"><nlm:affiliation>Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY 12180</wicri:regionArea>
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<term>Fluorescent Dyes</term>
<term>Guanosine Monophosphate (chemistry)</term>
<term>Molecular Sequence Data</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>ADN simple brin (analyse)</term>
<term>Colorants fluorescents</term>
<term>Composition en bases nucléiques</term>
<term>Données de séquences moléculaires</term>
<term>Guanosine monophosphate ()</term>
<term>Séquence nucléotidique</term>
<term>Électrochromatographie capillaire</term>
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<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ADN simple brin</term>
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<keywords scheme="MESH" xml:lang="en"><term>Base Composition</term>
<term>Base Sequence</term>
<term>Capillary Electrochromatography</term>
<term>Fluorescent Dyes</term>
<term>Molecular Sequence Data</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Colorants fluorescents</term>
<term>Composition en bases nucléiques</term>
<term>Données de séquences moléculaires</term>
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<front><div type="abstract" xml:lang="en">The separation of fluorescent-labeled ssDNA fragments of equal length based on differences in sequence was achieved through the use of guanosine gels (G-gels) formed by guanosine-5'-monophosphate (GMP) in capillary gel electrokinetic chromatography (CGEKC) with LIF detection. Baseline resolution was achieved for homodimers and homopentamers of A, T, and C. G-gel CGEKC provided better resolution than CZE, MEKC, or a sieving gel in CGE. Resolution improved with increasing GMP, indicating that the interaction is linked to structural organization of the G-gel. Fluorescence intensity and anisotropy show that the order of interaction with G-gels is T>C>A. We then investigated four conformationally similar, polymorphic 76-mers with A/G substitutions that are utilized in forensic DNA typing. Resolution was achieved by CGEKC but not CZE or CGE. In CGEKC, the negatively charged G-gels and oligonucleotides electromigrate toward the positive inlet while being driven by EOF to the negative outlet. The net forward velocity is the greatest for oligonucleotides most closely associated with the slower, more cumbersome G-gel network. For the 76-mers, resolution increases with increasing difference in guanosine content between strands and, for a given difference, with increasing total guanosines in the strands.</div>
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Serveur d'exploration MERS

Guanosine gel for sequence-dependent separation of polymorphic ssDNA.

Guanosine gel for sequence-dependent separation of polymorphic ssDNA.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki

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