Adaptor Ligation-Based Polymerase Chain Reaction-Mediated Walking
| Autor: | Linas Padegimas, Nancy A. Reichert |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
DNA
Plant Sequence analysis Restriction Mapping Biophysics Biology Genes Plant Polymerase Chain Reaction Zea mays Biochemistry DNA sequencing law.invention chemistry.chemical_compound law Promoter Regions Genetic Molecular Biology Polymerase chain reaction DNA Primers Exonuclease III Base Sequence Phosphogluconate Dehydrogenase Inverse polymerase chain reaction Sequence Analysis DNA Cell Biology Molecular biology genomic DNA Exodeoxyribonucleases Peroxidases chemistry biology.protein Nucleic Acid Conformation Indicators and Reagents Ligation DNA |
| Zdroj: | Analytical Biochemistry. 260:149-153 |
| ISSN: | 0003-2697 |
| DOI: | 10.1006/abio.1998.2719 |
| Popis: | An improved method of adaptor ligation PCR was developed for isolation of unknown sequences flanking a known DNA sequence. It was determined that the specificity of the adaptor ligation-based walking technique could be significantly enhanced by using uniquely blocked adaptors along with removal of unligated genomic DNA by exonuclease III digestion. This technique was utilized to isolate three novel promoter regions from three different Zea mays (maize) peroxidase genes. Sequences encoding a putative maize 6-phosphogluconate dehydrogenase gene were also isolated and confirmed by sequence analysis. The described improvements could be applied to other existing adaptor ligation-based PCR walking techniques. |
| Databáze: | OpenAIRE |
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