A simplified method for mapping deletion/substitution mutants of bacteriophage lambda
| Autor: | R.W. Davies, T.G. Fanning, P.H. Schreier, D.E. Büchel |
|---|---|
| Rok vydání: | 1977 |
| Předmět: |
Base pair
Mutant Biophysics EcoRI Biochemistry Coliphages chemistry.chemical_compound Escherichia coli Methods Molecular Biology Gel electrophoresis Nuclease Deoxyribonucleases biology Chromosome Mapping Cell Biology DNA Restriction Enzymes Molecular biology Molecular Weight Restriction enzyme chemistry DNA Viral Mutation biology.protein Electrophoresis Polyacrylamide Gel Chromosome Deletion DNA Heteroduplex |
| Zdroj: | Analytical biochemistry. 81(1) |
| ISSN: | 0003-2697 |
| Popis: | A convenient and rapid method for mapping deletion/substitution mutants of phage λ is presented. The method involves: (a) forming heteroduplex DNA molecules between the wild-type and the mutant: (b) digestion of the single-stranded regions with S1 nuclease; (c) cleavage of a portion of the remaining duplex DNA with EcoRI nuclease or any convenient restriction endonuclease; and (d) separation of the resulting DNA fragments by gel electrophoresis. Using three deletion/substitution mutants with known endpoints, we show that the values obtained by this method deviate, on the average, by ±120 base pairs from the values obtained by electron microscopy. |
| Databáze: | OpenAIRE |
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