<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>2007-1124</journal-id>
<journal-title><![CDATA[Revista mexicana de ciencias pecuarias]]></journal-title>
<abbrev-journal-title><![CDATA[Rev. mex. de cienc. pecuarias]]></abbrev-journal-title>
<issn>2007-1124</issn>
<publisher>
<publisher-name><![CDATA[Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S2007-11242016000400405</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Modificación del método de tiocianato de guanidina para extraer ADN de semen para análisis genómico en mamíferos]]></article-title>
<article-title xml:lang="en"><![CDATA[Modification of a guanidinium thiocyanate to extract DNA from semen for genomic analysis in mammals]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Alarcón-Zúñiga]]></surname>
<given-names><![CDATA[Baldomero]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Zepeda-Batista]]></surname>
<given-names><![CDATA[José Luis]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Ruíz-Flores]]></surname>
<given-names><![CDATA[Agustín]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Gómez-Meza]]></surname>
<given-names><![CDATA[Luis Joaquín]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[García-Muñiz]]></surname>
<given-names><![CDATA[José Guadalupe]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Núñez-Domínguez]]></surname>
<given-names><![CDATA[Rafael]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Ramírez-Valverde]]></surname>
<given-names><![CDATA[Rodolfo]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Villegas-Velázquez]]></surname>
<given-names><![CDATA[Itzel]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad Autónoma Chapingo Departamento de Zootecnia Posgrado en Producción Animal]]></institution>
<addr-line><![CDATA[Chapingo Estado de México]]></addr-line>
<country>Mexico</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2016</year>
</pub-date>
<volume>7</volume>
<numero>4</numero>
<fpage>405</fpage>
<lpage>413</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.org.mx/scielo.php?script=sci_arttext&amp;pid=S2007-11242016000400405&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.mx/scielo.php?script=sci_abstract&amp;pid=S2007-11242016000400405&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.mx/scielo.php?script=sci_pdf&amp;pid=S2007-11242016000400405&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen: Los análisis genómicos y transcriptómicos para selección y mejoramiento genético animal requieren ADN o ARN de alta concentración y pureza, proveniente de diferentes tejidos incluyendo semen. Los métodos usualmente utilizados para extraer ADN de semen son menos efectivos en cantidad y calidad de ADN, debido a los solventes y diluyente utilizados para la conservación, características físico-químicas de los espermatozoides, y fracción no celular del eyaculado. En este estudio, se proponen modificaciones al método de tiocianato de guanidina, incluyendo un segundo lavado de la muestra con solución buffer fosfato, y dos lavados con solventes orgánicos, uno fuerte (fenol:cloroformo:alcohol isoamílico) y uno débil (cloroformo:alcohol isoamílico), para retirar la proteína y diluyente presentes en la muestra. Además, se propone una incubación por separado con ARNasa para reducir contaminación de ácidos nucleicos en la medición y elaboración de diluciones para amplificación por PCR. La precipitación agregó al isopropanol de la metodología original 3 M de acetato de sodio para retirar restos de posibles inhibidores de la PCR. Finalmente, se incluyeron centrifugaciones de alta velocidad y decantaciones para evitar la necesidad de separación mecánica del ADN y la proteína. El ADN extraído con el método propuesto no presentó degradación, y la calidad y cantidad fueron mejores (P&lt;0.0001), encontrándose una media de 1.84±0.09 en el rango 260/280 y 156.99±7.29 ng/µl para la variable de concentración. El presente método de extracción es una alternativa de bajo costo, viable para obtener ADN de semen con características necesarias para análisis genómicos en mamíferos.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract: The genomic and transcriptomic analyses for selection and genetic improvement require DNA or RNA of high concentration and purity, coming from different tissues, including semen. The DNA may be from hair, saliva, cartilage, blood, or semen. The methods usually used to extract DNA from semen have low efficiency in terms of quantity and quality. This is due to the solvents and dilutors used for semen conservation, physical and chemical characteristics of sperms, and non-cellular fraction of ejaculate. In this study, modifications to the guanidinium thiocyanate method are proposed, including a second washing of the sample using a phosphate buffer solution, and two washing with organic solvents, one strong (phenol:chloroform:Isoamyl alcohol), and one weak (chloroform:isoamyl alcohol), to get rid of the protein and dilutors present in the sample. Additionally, it is proposed a separate incubation with RNA-ase to reduce contamination of nucleic acids during measurement and dilution preparation for PCR amplification. The precipitation added to the isopropanol of the original method 3 M sodium acetate to separate the residuals of potential PCR inhibitors. Finally, there were included high-speed centrifugation and decantation to avoid the need for mechanical separation of the DNA and protein. The DNA extracted with the modified method did not present degradation, and the quality and quantity were better (P&lt;0.0001) than the original, with a mean of 1.84±0.09 in the range of 260/ 280 and 156.99±7.29 ng/µl for concentration. The present method of extraction is a viable low-cost alternative to obtain DNA from semen with the necessary characteristics for genomic analysis in mammals.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[ADN genómico]]></kwd>
<kwd lng="es"><![CDATA[Extracción de ADN]]></kwd>
<kwd lng="es"><![CDATA[Semen]]></kwd>
<kwd lng="es"><![CDATA[Mamíferos]]></kwd>
<kwd lng="en"><![CDATA[Genomic DNA]]></kwd>
<kwd lng="en"><![CDATA[DNA extraction]]></kwd>
<kwd lng="en"><![CDATA[Semen]]></kwd>
<kwd lng="en"><![CDATA[Mammals]]></kwd>
</kwd-group>
</article-meta>
</front><back>
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