Determination of human alcohol dehydrogenase and acetaldehyde dehydrogenase genotypes by single strand conformation polymorphism in discontinuous buffer electrophoresis
| Autor: | Hans Türler, Olivier Georges Golaz, Denis F. Hochstrasser, Maristela Monteiro, Luc P. Balant, Claude Walzer, Jean-Pierre von Wartburg |
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| Rok vydání: | 1993 |
| Předmět: |
Genotype
Clinical Biochemistry Molecular Sequence Data DNA Single-Stranded Aldehyde Oxidoreductases / genetics Buffers Biochemistry Polymerase Chain Reaction Analytical Chemistry law.invention Silver stain ddc:616.89 chemistry.chemical_compound law ddc:570 DNA Single-Stranded / chemistry Humans Alcohol Dehydrogenase / genetics Polyacrylamide gel electrophoresis Polymerase chain reaction Alcohol dehydrogenase ddc:616 Polymorphism Genetic biology Base Sequence Oligonucleotide Alcohol Dehydrogenase Single-strand conformation polymorphism Aldehyde Oxidoreductases chemistry Electrophoresis Polyacrylamide Gel / methods biology.protein Nucleic Acid Conformation Electrophoresis Polyacrylamide Gel DNA |
| Zdroj: | Electrophoresis, Vol. 14, No 1 (1993) pp. 566-569 |
| ISSN: | 0173-0835 |
| Popis: | Under appropriate conditions single strand conformation polymorphism (SSCP) analysis of polymerase chain reaction (PCR) products allows the detection of single base mutations in a given DNA fragment. We adapted this method for the routine determination of allele variants of human alcohol and acetaldehyde dehydrogenase without radioisotopic labeling. After PCR amplification of the selected exon, the DNA fragments were heat‐denatured and loaded on a polyacrylamide gel containing glycerol. For electrophoresis a discontinuous buffer system was used with sulfate as leading ion and borate as trailing ion. The DNA bands were revealed by silver staining. Acrylamide concentrations, ionic strength and electrophoresis temperature were systematically investigated for each DNA fragment. The polymorphisms detected by SSCP were identical to those found by hybridization with 32P‐labeled allele‐specific oligonucleotides. This method avoids the use of radioactivity, is less expensive and simpler than the allele‐specific oligonucleotide (ASO) methodology and thus particularly suited for routine analysis. |
| Databáze: | OpenAIRE |
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