Estandarización de un protocolo sencillo para la extracción de ADN genómico de levaduras
<p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: #4d4d4d; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold; mso-ansi-language: EN-US;" lang="EN-US"...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | Spanish |
| Published: |
Universidad Nacional de Colombia
2009-10-01
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| Series: | Revista Colombiana de Biotecnología |
| Subjects: | |
| Online Access: | http://www.revistas.unal.edu.co/index.php/biotecnologia/article/view/10339 |
| _version_ | 1818496596194623488 |
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| author | Esteban Osorio-Cadavid Mauricio Ramírez William Andrés López Luz Adriana Mambuscay |
| author_facet | Esteban Osorio-Cadavid Mauricio Ramírez William Andrés López Luz Adriana Mambuscay |
| author_sort | Esteban Osorio-Cadavid |
| collection | DOAJ |
| description | <p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: #4d4d4d; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold; mso-ansi-language: EN-US;" lang="EN-US"><span style="font-family: Times New Roman;">Standardising a simple protocol for extracting yeast from genomic DNA</span></span></strong></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: black; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold; mso-ansi-language: EN-US;" lang="EN-US"><span style="font-family: Times New Roman;"> </span></span></strong></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: black; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold;"><span style="font-family: Times New Roman;">Resumen: </span></span></strong><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond;">Se estandarizó un protocolo rápido, sencillo y de bajo costo para la extracción de ADN genómico de levaduras a partir de lisis de la pared celular mediante tratamiento enzimático y precipitación por alcoholes. El empleo de la enzima Beta-glucoronidasa en reemplazo de la enzima Zimolasa, permitió obtener ADN en alta concentración (124,9±30,2 ng/μl) y de buena calidad (A260/A280 nm =1,86±0,1), ideal para su uso en estudios de biología molecular. Además, se adicionó un paso de incubación del ADN obtenido a 100° C para inactivar ADNasas. La calidad del ADN obtenido fue evaluada por medio de la amplificación de la región ITS1-5.8S-ITS2, presentando bandas definidas y cuantificables (entre 380 y 880 pb) ideales para estudios de identificación molecular y filogenia.</span></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: black; font-family: Garamond-Bold; mso-bidi-font-family: Garamond-Bold;"><span style="font-family: Times New Roman;">Palabras clave: </span></span></strong><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond;">levadura; extracción de ADN; Beta-Glucoronidasa.</span></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"> </p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold; mso-ansi-language: EN-US;" lang="EN-US"><span style="font-family: Times New Roman;">Abstract: </span></span></strong><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond;">A quick, simple and low-cost protocol for extracting genomic DNA from yeast by cell wall lysis involving enzymatic treatment and alcoholic precipitation was standardised. Higher DNA yields (124.9±30.2 ng/μl) were obtained by using beta-glucuronidase instead of zymolyase; these had very high quality (A260/A280 nm = 1.86±0.1) and would be suitable for use in molecular biology assays. Moreover, a DNAse inactivation step was also introduced by incubation at 100 °C to further ensure DNA stability. </span><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond; mso-ansi-language: EN-US;" lang="EN-US">DNA quality was assayed by PCR amplification </span><span style="font-size: 10pt; font-family: Garamond; mso-bidi-font-family: Garamond; mso-ansi-language: EN-US;" lang="EN-US">of the ITS1-5.8S-ITS2 region, revealing defined, quantifiable 380 to 880 bp bands. These results show that the protocol is ideal for molecular identification and phylogenetic studies.</span><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond;"></span></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; font-family: Garamond-Bold; mso-bidi-font-family: Garamond-Bold; mso-ansi-language: EN-US;" lang="EN-US"><span style="font-family: Times New Roman;">Key words: </span></span></strong><span style="font-size: 10pt; font-family: Garamond; mso-bidi-font-family: Garamond; mso-ansi-language: EN-US;" lang="EN-US">Yeast; DNA extraction; beta-glucoronidase.</span></p> |
| first_indexed | 2024-12-10T18:35:01Z |
| format | Article |
| id | doaj.art-d7337e38f379434aa5d0ec7240c4b60a |
| institution | Directory Open Access Journal |
| issn | 0123-3475 1909-8758 |
| language | Spanish |
| last_indexed | 2024-12-10T18:35:01Z |
| publishDate | 2009-10-01 |
| publisher | Universidad Nacional de Colombia |
| record_format | Article |
| series | Revista Colombiana de Biotecnología |
| spelling | doaj.art-d7337e38f379434aa5d0ec7240c4b60a2022-12-22T01:37:50ZspaUniversidad Nacional de ColombiaRevista Colombiana de Biotecnología0123-34751909-87582009-10-011111251319786Estandarización de un protocolo sencillo para la extracción de ADN genómico de levadurasEsteban Osorio-Cadavid0Mauricio Ramírez1William Andrés López2Luz Adriana Mambuscay3InvestigadorInvestigadorInvestigadorInvestigador<p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: #4d4d4d; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold; mso-ansi-language: EN-US;" lang="EN-US"><span style="font-family: Times New Roman;">Standardising a simple protocol for extracting yeast from genomic DNA</span></span></strong></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: black; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold; mso-ansi-language: EN-US;" lang="EN-US"><span style="font-family: Times New Roman;"> </span></span></strong></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: black; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold;"><span style="font-family: Times New Roman;">Resumen: </span></span></strong><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond;">Se estandarizó un protocolo rápido, sencillo y de bajo costo para la extracción de ADN genómico de levaduras a partir de lisis de la pared celular mediante tratamiento enzimático y precipitación por alcoholes. El empleo de la enzima Beta-glucoronidasa en reemplazo de la enzima Zimolasa, permitió obtener ADN en alta concentración (124,9±30,2 ng/μl) y de buena calidad (A260/A280 nm =1,86±0,1), ideal para su uso en estudios de biología molecular. Además, se adicionó un paso de incubación del ADN obtenido a 100° C para inactivar ADNasas. La calidad del ADN obtenido fue evaluada por medio de la amplificación de la región ITS1-5.8S-ITS2, presentando bandas definidas y cuantificables (entre 380 y 880 pb) ideales para estudios de identificación molecular y filogenia.</span></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; color: black; font-family: Garamond-Bold; mso-bidi-font-family: Garamond-Bold;"><span style="font-family: Times New Roman;">Palabras clave: </span></span></strong><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond;">levadura; extracción de ADN; Beta-Glucoronidasa.</span></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"> </p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; font-family: AGaramondPro-Bold; mso-bidi-font-family: AGaramondPro-Bold; mso-ansi-language: EN-US;" lang="EN-US"><span style="font-family: Times New Roman;">Abstract: </span></span></strong><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond;">A quick, simple and low-cost protocol for extracting genomic DNA from yeast by cell wall lysis involving enzymatic treatment and alcoholic precipitation was standardised. Higher DNA yields (124.9±30.2 ng/μl) were obtained by using beta-glucuronidase instead of zymolyase; these had very high quality (A260/A280 nm = 1.86±0.1) and would be suitable for use in molecular biology assays. Moreover, a DNAse inactivation step was also introduced by incubation at 100 °C to further ensure DNA stability. </span><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond; mso-ansi-language: EN-US;" lang="EN-US">DNA quality was assayed by PCR amplification </span><span style="font-size: 10pt; font-family: Garamond; mso-bidi-font-family: Garamond; mso-ansi-language: EN-US;" lang="EN-US">of the ITS1-5.8S-ITS2 region, revealing defined, quantifiable 380 to 880 bp bands. These results show that the protocol is ideal for molecular identification and phylogenetic studies.</span><span style="font-size: 10pt; color: black; font-family: Garamond; mso-bidi-font-family: Garamond;"></span></p><p class="MsoNormal" style="margin: 0cm 0cm 0pt; mso-layout-grid-align: none;"><strong><span style="font-size: 10pt; font-family: Garamond-Bold; mso-bidi-font-family: Garamond-Bold; mso-ansi-language: EN-US;" lang="EN-US"><span style="font-family: Times New Roman;">Key words: </span></span></strong><span style="font-size: 10pt; font-family: Garamond; mso-bidi-font-family: Garamond; mso-ansi-language: EN-US;" lang="EN-US">Yeast; DNA extraction; beta-glucoronidase.</span></p>http://www.revistas.unal.edu.co/index.php/biotecnologia/article/view/10339levaduraextracción de ADNBeta-GlucoronidasaYeastDNA extractionbeta-glucoronidase |
| spellingShingle | Esteban Osorio-Cadavid Mauricio Ramírez William Andrés López Luz Adriana Mambuscay Estandarización de un protocolo sencillo para la extracción de ADN genómico de levaduras Revista Colombiana de Biotecnología levadura extracción de ADN Beta-Glucoronidasa Yeast DNA extraction beta-glucoronidase |
| title | Estandarización de un protocolo sencillo para la extracción de ADN genómico de levaduras |
| title_full | Estandarización de un protocolo sencillo para la extracción de ADN genómico de levaduras |
| title_fullStr | Estandarización de un protocolo sencillo para la extracción de ADN genómico de levaduras |
| title_full_unstemmed | Estandarización de un protocolo sencillo para la extracción de ADN genómico de levaduras |
| title_short | Estandarización de un protocolo sencillo para la extracción de ADN genómico de levaduras |
| title_sort | estandarizacion de un protocolo sencillo para la extraccion de adn genomico de levaduras |
| topic | levadura extracción de ADN Beta-Glucoronidasa Yeast DNA extraction beta-glucoronidase |
| url | http://www.revistas.unal.edu.co/index.php/biotecnologia/article/view/10339 |
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