A chemical method for introducing haptens onto DNA probes

Keller, George H.; Cumming, Cecilia U.; Huang, Dao-Pei; Manak, Mark M.; Ting, R. C.

doi:10.1016/0003-2697(88)90656-2

Cited by 64 publications

(15 citation statements)

References 18 publications

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“…Serum DNA was extracted using a standard protocol [18]. In brief, 250 µl of serum was digested with 250 µg/ml proteinase K (Sigma, St Louis, MO) in 150 m m NaCl, 10 m m EDTA, 10 m m Tris–HCl (pH 7·6), 2% sodium dodecyl sulphate (SDS) and 5 µg/ml calf thymus DNA (Sigma), overnight at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

High frequency of hepatitis B virus infection in patients with β‐thalassemia receiving multiple transfusions

Singh

Pradhan²,

Singh³

et al. 2003

Vox Sanguinis

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Section: Methodsmentioning

confidence: 99%

High frequency of hepatitis B virus infection in patients with β‐thalassemia receiving multiple transfusions

Singh

Pradhan²,

Singh³

et al. 2003

Vox Sanguinis

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“…The chemical modification of guanine residues in DNA at NH9-functions has also been described^3 81 . A more randomly labeling of DNA has been achieved by bromination and subsequent reaction with a hapten at C-8 of purines, C-5 of cytosine and possibly C-6 of thymine [39] .…”

Section: Discussionmentioning

confidence: 99%

Non-radioactive Labeling and Detection of Nucleic Acids. IV. Synthesis and Properties of Digoxigenin-modified 2´ -Deoxyuridine-5’ -triphosphates and a Photoactivatable Analog of Digoxigenin (Photodigoxigenin)

Mühlegger

Huber²,

Eltz³

et al. 1990

Biological Chemistry Hoppe-Seyler

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The chemical syntheses of novel digoxigenin-derivatized compounds are described which are modified substrates for enzymatically or photochemically non-radioactive digoxigenin labeling of nucleic acids. Various activated digoxigenin-haptens are coupled to 5-aminoallyl-substituted 2'-deoxyuridine-5'-triphosphate. This results in digoxigeninmodified nucleoside triphosphates of variable spacer lengths (Dig-[4]-dUTP/Dig-[ll]-dUTP/Dig-[16]-dUTP) which can be used as substrates for enzymatic labeling of DNA with digoxigenin-haptens by Klenow enzyme-catalysed random-primed synthesis. In addition the synthesis of N-, a photoactivatable analog of digoxigenin, is described which can be applied for photolabeling of DNA and RNA with digoxigenin-haptens leaving the nucleic acid molecules intact. Nicht-radioaktiveMarkierung und Detektion von Nucleinsäuren: IV. Synthese und Eigenschaften von Digoxigenin-modifizierten 2'-Desoxyuridin-5'-triphosphaten und eines photoaktivierbaren Analogons (Photodigoxigenin) Zusammenfassung: Es werden die chemischen Synthesen von solchen neuen, Digoxigenin-derivatisierten Komponenten beschrieben, die als modifizierte Substrate zur enzymatischen oder photochemischen nicht-radioaktiven Digoxigenin-Markierung von Nucleinsäuren eingesetzt werden können. Verschiedene aktivierte Digoxigenin-Haptene werden an 5-Aminoallyl-substituierte 2'-Desoxyuridin-5'-triphosphate gekoppelt. Es resultieren Digoxigenin-modifizierte Nucleosidtriphosphate mit unterschiedlichen Spacerlängen (Dig-[4]-dUTP/Dig-[ll]-dUTP/Dig-[16]-dUTP), die als Substrate zur enzymatischen Markierung von DNA mit Digoxigenin-Haptenen über die Klenow-Enzym-katalysierte Random-primed-Synthese eingesetzt werden können. Zusätzlich wird die Synthese von yV-[4]-Azidobenzoyl]-;V-[3-O-digoxigenininyl)methylcarbonyl]-l,8-diamino-3,6-dioxaoctan (Photodigoxigenin), einem photoaktivierbaren Analogon von Digoxigenin, beschrieben. Dieses Substrat kann zur Photomarkierung von DNA und RNA mit Digoxigenin-Haptenen angewendet werden; bei dieser Markierungsreaktion bleiben die Nucleinsäu-remoleküle intakt.

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“…The preparation of DNA from both tissue cells and blood often involve lengthy purification procedures (Maniatis et aI., 1982;Keller et aL, 1988;Laird et al, 1991).…”

Section: In~oducfionmentioning

confidence: 99%

A simple method of DNA extraction from whole tissues and blood using glass powder for detection of transgenic animals by PCR

Attal

Cajero-Juarez

Houdebiné

1995

Transgenic Research

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A chemical method for introducing haptens onto DNA probes

Cited by 64 publications

References 18 publications

High frequency of hepatitis B virus infection in patients with β‐thalassemia receiving multiple transfusions

High frequency of hepatitis B virus infection in patients with β‐thalassemia receiving multiple transfusions

Non-radioactive Labeling and Detection of Nucleic Acids. IV. Synthesis and Properties of Digoxigenin-modified 2´ -Deoxyuridine-5’ -triphosphates and a Photoactivatable Analog of Digoxigenin (Photodigoxigenin)

A simple method of DNA extraction from whole tissues and blood using glass powder for detection of transgenic animals by PCR

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