Peptide nucleic acid pre-gel hybridization: An alternative to Southern hybridization
| Autor: | James M. Coull, Michael Egholm, Martin Fuchs, Xian-Wei Yao, Heather Perry-O’keefe |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1996 |
| Předmět: |
Gel electrophoresis
Multidisciplinary Chromatography Gel electrophoresis of nucleic acids Peptide nucleic acid Hybridization probe musculoskeletal neural and ocular physiology Molecular Probe Techniques Nucleic Acid Hybridization Biological Sciences Electrophoresis chemistry.chemical_compound Blotting Southern Capillary electrophoresis chemistry Biochemistry biological sciences cardiovascular system Peptides tissues DNA Southern blot |
| Popis: | We have found that it is possible to use labeled peptide nucleic acid (PNA)-oligomers as probes in pre-gel hybridization experiments, as an alternative for Southern hybridization. In this technique, the PNA probe is hybridized to a denatured DNA sample at low ionic strength and the mixture is loaded directly on to an electrophoresis system for size separation. Ensuing gel electrophoresis separates the single-stranded DNA fragments by length. The neutral backbone of PNA allows for hybridization at low ionic strength and imparts very low mobility to excess PNA. Detection of the bound PNA is possible by direct fluorescence detection with capillary electrophoresis, or the DNA/PNA hybrids can be blotted onto a membrane and detected with standard chemiluminescent techniques. Efficient single bp discrimination was achieved routinely using both capillary and slab-gel electrophoresis. |
| Databáze: | OpenAIRE |
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