MersV1, Main, Exploration, bibRecord, 004515

Oligodeoxynucleotides capable of binding to GC-rich regions in DNA are inappropriately synthesized during random hexanucleotide-primed labeling reactions.

Identifieur interne : 004515 ( Main/Exploration ); précédent : 004514; suivant : 004516

Oligodeoxynucleotides capable of binding to GC-rich regions in DNA are inappropriately synthesized during random hexanucleotide-primed labeling reactions.

Auteurs : J H Chen [Canada] ; H C Birnboim

Source :

DNA and cell biology [ 1044-5498 ] ; 1993.

RBID : pubmed:8471165

Descripteurs français

KwdFr :
- ADN simple brin (analyse), Artéfacts, Données de séquences moléculaires, Marquage isotopique, Sondes d'ADN, Séquence nucléotidique, Technique de Northern (), Technique de Southern ().
MESH :
- analyse : ADN simple brin.
- Artéfacts, Données de séquences moléculaires, Marquage isotopique, Sondes d'ADN, Séquence nucléotidique, Technique de Northern, Technique de Southern.

English descriptors

KwdEn :
- Artifacts, Base Sequence, Blotting, Northern (methods), Blotting, Southern (methods), DNA Probes, DNA, Single-Stranded (analysis), Isotope Labeling, Molecular Sequence Data.
MESH :
- chemical , analysis : DNA, Single-Stranded.
- chemical : DNA Probes.
- methods : Blotting, Northern, Blotting, Southern.
- Artifacts, Base Sequence, Isotope Labeling, Molecular Sequence Data.

Abstract

Random hexanucleotide labeling is a commonly used and powerful technique for preparing radiolabeled nucleic acid probes. Because of the dependence of DNA polymerase upon single-stranded DNA as a template, the technique should theoretically also be useful for detecting small amounts of single-stranded DNA in the presence of a larger amount of double-stranded DNA. In the course of such an experiment, we identified an unexpected reaction that could contribute to background problems often encountered during Southern and Northern blotting. In the complete absence of template, random hexanucleotides appear capable of functioning both as template as well as primer, giving rise to labeled oligonucleotides as large as 20-mers. Some of these bind very tightly to GC-rich regions in target DNA immobilized on membranes, resisting high-stringency washing conditions. By choosing shorter incubation times and including sufficient single-stranded template, the problem may be minimized.

DOI: 10.1089/dna.1993.12.177
PubMed: 8471165

Affiliations:

Canada

Le document en format XML

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<term>Isotope Labeling</term>
<term>Molecular Sequence Data</term>
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<term>Données de séquences moléculaires</term>
<term>Marquage isotopique</term>
<term>Sondes d'ADN</term>
<term>Séquence nucléotidique</term>
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<front><div type="abstract" xml:lang="en">Random hexanucleotide labeling is a commonly used and powerful technique for preparing radiolabeled nucleic acid probes. Because of the dependence of DNA polymerase upon single-stranded DNA as a template, the technique should theoretically also be useful for detecting small amounts of single-stranded DNA in the presence of a larger amount of double-stranded DNA. In the course of such an experiment, we identified an unexpected reaction that could contribute to background problems often encountered during Southern and Northern blotting. In the complete absence of template, random hexanucleotides appear capable of functioning both as template as well as primer, giving rise to labeled oligonucleotides as large as 20-mers. Some of these bind very tightly to GC-rich regions in target DNA immobilized on membranes, resisting high-stringency washing conditions. By choosing shorter incubation times and including sufficient single-stranded template, the problem may be minimized.</div>
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Serveur d'exploration MERS

Oligodeoxynucleotides capable of binding to GC-rich regions in DNA are inappropriately synthesized during random hexanucleotide-primed labeling reactions.

Oligodeoxynucleotides capable of binding to GC-rich regions in DNA are inappropriately synthesized during random hexanucleotide-primed labeling reactions.

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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