Comparison of two detection systems to reveal AFLP markers in plants
Since their development, AFLP (amplified fragment length polymorphism) markers have been used for a wide variety of analyses and, up to this day, are considered highly informative, robust, and reproducible molecular markers. Originally, the visualization of the amplified fragments was done in polyac...
| Autores principales: | , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
2018
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/2664 http://www.nrcresearchpress.com/doi/10.1139/cjb-2014-0047#.WyuX_jdKjcs https://doi.org/10.1139/cjb-2014-0047 |
| _version_ | 1855035004130689024 |
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| author | Cara, Nicolás Marfil, Carlos Federico Garcia Lampasona, Sandra Claudia Masuelli, Ricardo Williams |
| author_browse | Cara, Nicolás Garcia Lampasona, Sandra Claudia Marfil, Carlos Federico Masuelli, Ricardo Williams |
| author_facet | Cara, Nicolás Marfil, Carlos Federico Garcia Lampasona, Sandra Claudia Masuelli, Ricardo Williams |
| author_sort | Cara, Nicolás |
| collection | INTA Digital |
| description | Since their development, AFLP (amplified fragment length polymorphism) markers have been used for a wide variety of analyses and, up to this day, are considered highly informative, robust, and reproducible molecular markers. Originally, the visualization of the amplified fragments was done in polyacrylamide gels, followed by silver staining or by developing in an X-ray plate, when radioactivity is used. In the last 14 years, capillary electrophoresis of fluorescently labeled fragments has been gradually replacing gel-based systems. However, the latter continue to be better for isolating and cloning AFLP fragments. In this report, we compare the results obtained by capillary electrophoresis with those from silver staining. We found that if fluorescence-labeled amplification products are loaded in a polyacrylamide gel, duplicated bands (doublets) are seen. This phenomenon is probably due to a delay in the migration of the strand that carries the fluorophore. Therefore, we recommend a minimum separation of 4 bp from the nearest fragment to the target fragment for its unambiguous identification and isolation. If this requirement is not fulfilled, an alternative is to make new amplifications using the same primer combination, but with unlabeled primers. |
| format | info:ar-repo/semantics/artículo |
| id | INTA2664 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| record_format | dspace |
| spelling | INTA26642023-12-21T16:31:04Z Comparison of two detection systems to reveal AFLP markers in plants Cara, Nicolás Marfil, Carlos Federico Garcia Lampasona, Sandra Claudia Masuelli, Ricardo Williams Marcadores Genéticos Polimorfismo de Longitud de Fragmentos Amplificados Plantas Electrofóresis Gel Poliacrilamida Genetic Markers Amplified Fragment Length Polymorphism Plants Polyacrylamide Gel Electrophoresis AFLP Marcadores Moleculares Since their development, AFLP (amplified fragment length polymorphism) markers have been used for a wide variety of analyses and, up to this day, are considered highly informative, robust, and reproducible molecular markers. Originally, the visualization of the amplified fragments was done in polyacrylamide gels, followed by silver staining or by developing in an X-ray plate, when radioactivity is used. In the last 14 years, capillary electrophoresis of fluorescently labeled fragments has been gradually replacing gel-based systems. However, the latter continue to be better for isolating and cloning AFLP fragments. In this report, we compare the results obtained by capillary electrophoresis with those from silver staining. We found that if fluorescence-labeled amplification products are loaded in a polyacrylamide gel, duplicated bands (doublets) are seen. This phenomenon is probably due to a delay in the migration of the strand that carries the fluorophore. Therefore, we recommend a minimum separation of 4 bp from the nearest fragment to the target fragment for its unambiguous identification and isolation. If this requirement is not fulfilled, an alternative is to make new amplifications using the same primer combination, but with unlabeled primers. EEA La Consulta Fil: Cara, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; Argentina Fil: Marfil, Carlos Fedrico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; Argentina Fil: Garcia Lampasona, Sandra Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria La Consulta; Argentina Fil: Masuelli, Ricardo Williams. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria La Consulta; Argentina 2018-06-21T12:33:45Z 2018-06-21T12:33:45Z 2014-06 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/2664 http://www.nrcresearchpress.com/doi/10.1139/cjb-2014-0047#.WyuX_jdKjcs 1916-2790 1916-2804 https://doi.org/10.1139/cjb-2014-0047 eng info:eu-repo/semantics/restrictedAccess application/pdf Botany 92 (8) : 607-610 (2014) |
| spellingShingle | Marcadores Genéticos Polimorfismo de Longitud de Fragmentos Amplificados Plantas Electrofóresis Gel Poliacrilamida Genetic Markers Amplified Fragment Length Polymorphism Plants Polyacrylamide Gel Electrophoresis AFLP Marcadores Moleculares Cara, Nicolás Marfil, Carlos Federico Garcia Lampasona, Sandra Claudia Masuelli, Ricardo Williams Comparison of two detection systems to reveal AFLP markers in plants |
| title | Comparison of two detection systems to reveal AFLP markers in plants |
| title_full | Comparison of two detection systems to reveal AFLP markers in plants |
| title_fullStr | Comparison of two detection systems to reveal AFLP markers in plants |
| title_full_unstemmed | Comparison of two detection systems to reveal AFLP markers in plants |
| title_short | Comparison of two detection systems to reveal AFLP markers in plants |
| title_sort | comparison of two detection systems to reveal aflp markers in plants |
| topic | Marcadores Genéticos Polimorfismo de Longitud de Fragmentos Amplificados Plantas Electrofóresis Gel Poliacrilamida Genetic Markers Amplified Fragment Length Polymorphism Plants Polyacrylamide Gel Electrophoresis AFLP Marcadores Moleculares |
| url | http://hdl.handle.net/20.500.12123/2664 http://www.nrcresearchpress.com/doi/10.1139/cjb-2014-0047#.WyuX_jdKjcs https://doi.org/10.1139/cjb-2014-0047 |
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