Molecular Hybridization Probes Prepared with 4-Aminooxybutylamine
| Autor: | and Anna V. Kiseliova, Sergey M. Kalachikov, Vyacheslav A. Adarichev, Grigory M. Dymshits |
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| Rok vydání: | 1998 |
| Předmět: |
Transamination
Biomedical Engineering Biotin Pharmaceutical Science Bioengineering Butylamines Mice chemistry.chemical_compound medicine Animals Cells Cultured In Situ Hybridization Fluorescence Fluorescent Dyes Pharmacology Detection limit chemistry.chemical_classification medicine.diagnostic_test Aminooxybutylamine Hybridization probe Organic Chemistry DNA Combinatorial chemistry Molecular hybridization chemistry Mink Fluorescein Indicators and Reagents DNA Probes Biotechnology Fluorescence in situ hybridization |
| Zdroj: | Bioconjugate Chemistry. 9:671-675 |
| ISSN: | 1520-4812 1043-1802 |
| Popis: | A versatile method is described for preparing nonradioactive DNA probes for molecular hybridization. The method is based on the transamination reaction of double-stranded DNA with 4-aminooxybutylamine (ABA). To optimize the procedure for obtaining stable and sensitive hybridization probes, time of modification, pH, and reaction temperature were varied. The optimal reaction conditions allowed the preparation of nonradioactive-labeled DNA probes that met demands for optimal length, modification degree, stability, and sensitivity. The use of 4-aminooxybutylamine as a bifunctional reagent for DNA modification allowed the possibility of choosing an appropriate reporter group: biotin or one of the fluorochromes. For probes carrying biotin, a high specificity and high sensitivity detection limit of 1 pg of target DNA were demonstrated. In addition, the applicability of probes carrying fluorochromes for multicolor direct fluorescence in situ hybridization was described. |
| Databáze: | OpenAIRE |
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