A new sensitive non-isotopic method using sulfonated probes to detect picogram quantities of specific DNA sequences on blot hybridization
| Autor: | Driss Squalli, Martine Duchenne, Martine Joannes, Philippe Pouletty, Philippe Lebacq |
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| Rok vydání: | 1988 |
| Předmět: |
Chromatography
Resolution (mass spectrometry) Base Sequence Hybridization probe Immunochemistry Biophysics Nucleic Acid Hybridization DNA Biology Biochemistry Molecular biology DNA sequencing Nuclear DNA chemistry.chemical_compound chemistry Protein Biosynthesis Sulfones Restriction fragment length polymorphism Nitrocellulose Southern blot |
| Zdroj: | Journal of biochemical and biophysical methods. 15(5) |
| ISSN: | 0165-022X |
| Popis: | The use of non-radioactive systems to detect target DNA or RNA displays many advantages such as safe manipulation, potential use in non-specialized scientific area and prolonged lifetime of the probes (one year or more). We here describe a method we have improved and optimized using sulfonated DNA probes for hybridization on dot and Southern blots. Sulfonation is an easy chemical modification procedure which does not require enzymatic coctain as does nick-translation. Sensitivity of this method has been particularly improved by using a new blocking solution, containing heparin, which allows easy and fast detection of picogram quantities of DNa. This method allows the use of nitrocellulose as well as nylon membranes with very low background. Equal resolution is obtained in comparative experiments involving both sulfonated and 32 P-radiolabelled probes. Single copy gene sequences are readily detected in nuclear DNA. These results allow the use of this procedure for restriction fragment length polymorphism (RFLP) studies. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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