Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays

The extraction of high-quality DNA is essential for studies conducted at the molecular level in species with an abundance of contaminants in their tissues, such as some landraces of potato and maize, in which it is difficult to extract good-quality genomic DNA. Compounds such as polyphenols interfer...

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Autores principales:	Barra,Macarena, Salazar,Erika, Beltrán,María, Sagredo,Boris
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Chile. Facultad de Agronomía e Ingeniería Forestal 2012
Materias:	Activated charcoal DNA extraction genomic DNA polyphenols
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0718-16202012000300019
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spelling	oai:scielo:S0718-162020120003000192014-03-10Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea maysBarra,MacarenaSalazar,ErikaBeltrán,MaríaSagredo,Boris Activated charcoal DNA extraction genomic DNA polyphenols The extraction of high-quality DNA is essential for studies conducted at the molecular level in species with an abundance of contaminants in their tissues, such as some landraces of potato and maize, in which it is difficult to extract good-quality genomic DNA. Compounds such as polyphenols interfere with the amplification of DNA during polymerase chain reaction (PCR), which makes it difficult to conduct PCR-based studies ofmolecular markers. This article describes a simple and robust protocol for DNA isolation that was applied to potato and maize landraces and resulted in a high DNA concentration with excellent purity and quality. This method is based on the method of Krizman et al. (2006) with some modifications and was tested on potato leaves and young maize leaves. The protocol was compared with one commercial DNA extraction kit and three methods that were previously described for plant DNA extraction: Keb-Llanes et al. (2002), for plants with high polyphenolic content; Fulton et al. (1995), which was developed for Solanaceae and other herbaceous plants; and the original Krizman et al. (2006) cetyltrimethylammonium bromide (CTAB)-activated charcoal method . The method proposed is suitable for isolating DNA Irom potato and maize genotypes with high polyphenol contents, yielding high-quality DNA that can be utilized for molecular techniques that involve PCR, cutting with restriction enzymes and other applications. The method is short, similar to a commercial kit, but cheaper. The amount of DNA extracted from maize using our method is superior compared with other published methods tested in this article.info:eu-repo/semantics/openAccessPontificia Universidad Católica de Chile. Facultad de Agronomía e Ingeniería ForestalCiencia e investigación agraria v.39 n.3 20122012-12-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0718-16202012000300019en10.4067/S0718-16202012000300019
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Activated charcoal DNA extraction genomic DNA polyphenols
spellingShingle	Activated charcoal DNA extraction genomic DNA polyphenols Barra,Macarena Salazar,Erika Beltrán,María Sagredo,Boris Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays
description	The extraction of high-quality DNA is essential for studies conducted at the molecular level in species with an abundance of contaminants in their tissues, such as some landraces of potato and maize, in which it is difficult to extract good-quality genomic DNA. Compounds such as polyphenols interfere with the amplification of DNA during polymerase chain reaction (PCR), which makes it difficult to conduct PCR-based studies ofmolecular markers. This article describes a simple and robust protocol for DNA isolation that was applied to potato and maize landraces and resulted in a high DNA concentration with excellent purity and quality. This method is based on the method of Krizman et al. (2006) with some modifications and was tested on potato leaves and young maize leaves. The protocol was compared with one commercial DNA extraction kit and three methods that were previously described for plant DNA extraction: Keb-Llanes et al. (2002), for plants with high polyphenolic content; Fulton et al. (1995), which was developed for Solanaceae and other herbaceous plants; and the original Krizman et al. (2006) cetyltrimethylammonium bromide (CTAB)-activated charcoal method . The method proposed is suitable for isolating DNA Irom potato and maize genotypes with high polyphenol contents, yielding high-quality DNA that can be utilized for molecular techniques that involve PCR, cutting with restriction enzymes and other applications. The method is short, similar to a commercial kit, but cheaper. The amount of DNA extracted from maize using our method is superior compared with other published methods tested in this article.
author	Barra,Macarena Salazar,Erika Beltrán,María Sagredo,Boris
author_facet	Barra,Macarena Salazar,Erika Beltrán,María Sagredo,Boris
author_sort	Barra,Macarena
title	Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays
title_short	Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays
title_full	Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays
title_fullStr	Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays
title_full_unstemmed	Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays
title_sort	simple and robust dna extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof solanum tuberosum and zea mays
publisher	Pontificia Universidad Católica de Chile. Facultad de Agronomía e Ingeniería Forestal
publishDate	2012
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0718-16202012000300019
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Simple and robust DNA extraction method for the large-scale analysis of genotypes containing high polyphenolic content, such as landracesof Solanum tuberosum and Zea mays

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