An optimized protocol for the production of interdelta markers in Saccharomyces cerevisiae by using capillary electrophoresis
| Autor: | Francesco Grieco, Mariana Tristezza, Carmela Gerardi, Antonio F. Logrieco |
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| Rok vydání: | 2009 |
| Předmět: |
Microbiology (medical)
Genetic Markers Chromatography biology Staining and Labeling Saccharomyces cerevisiae Electrophoresis Capillary Nucleic acid amplification technique biology.organism_classification Microbiology Molecular biology Fluorescence DNA Fingerprinting Interdelta Capillary electrophoresis DNA profiling Delta Elements Genetic marker DNA Fungal Mycological Typing Techniques Molecular Biology Nucleic Acid Amplification Techniques |
| Zdroj: | Journal of microbiological methods 78 (2009): 286–291. doi:10.1016/j.mimet.2009.06.012 info:cnr-pdr/source/autori:Tristezza M., Gerardi C., Logrieco A. and Grieco F.*/titolo:An optimized protocol for the production of interdelta markers in Saccharomyces cerevisiae by using capillary electrophoresis./doi:10.1016%2Fj.mimet.2009.06.012/rivista:Journal of microbiological methods/anno:2009/pagina_da:286/pagina_a:291/intervallo_pagine:286–291/volume:78 |
| ISSN: | 1872-8359 |
| DOI: | 10.1016/j.mimet.2009.06.012 |
| Popis: | The amplification of genomic sequence blocks flanked by delta elements of retrotransposon origin has proved to be a very convenient method for molecular characterization of Saccharomyces cerevisiae strains. Fluorescent automated capillary electrophoresis (CE) was used to detect interdelta marker (IDM) patterns in S. cerevisiae, using the ABI Prism 3130 Genetic Analyzer. Main experimental parameters were studied and the optimal conditions for IDM amplification and samples run on the CE apparatus were determined. Fingerprints from fluorescent-labelled IDM produced using CE with the same sample analyzed by agarose electrophoresis (AE) were compared. The CE analysis was able to distinguish 43 different IDM profiles among 45 S. cerevisiae isolates with a discriminating capacity of 99.8%, whereas the AE analysis of the same samples allowed the identification of 27 different patterns (discriminatory power equal to 96%). Detection of fluorescent IDM was fast and reliable, and it facilitated data comparison. For the first time in our knowledge, the fluorescent CE proved to be well suited for IDM fingerprinting. Moreover, it could be routinely applied for the molecular differentiation of S. cerevisiae strains. |
| Databáze: | OpenAIRE |
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