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Comparación de dos kits de RT-PCR en la detección de ARNm de dos genes endógenos de papa (Solanum tuberosum spp. Andígena)
Keywords:
Solanum tuberosum, cox, actina, RT-PCR, papa, actin, potato (es)Downloads
RT-PCR es una técnica en la que usando ARN mensajero como molde, se obtiene complementario o cADN por transcripción inversa, y luego se amplifica uno de los cADN por PCR, mediante el uso de primers específicos. Esta técnica permite realizar estudios de expresión, a nivel de ARN mensajero. Con el propósito de implementar la técnica en papa (Solanum tuberosum spp. Andígena), se utilizaron plántulas cultivadas in vitro de la variedad Pastusa Suprema. Inicialmente se establecieron las condiciones para la extracción de ARN total usando el kit TRizol® Reagent de InvitrogenTM, con el que se obtuvieron excelentes resultados. Este ARN se usó como molde para evaluar dos kits: “ONE Step superScriptTM†y “SuperScriptâ„¢ First Strand Syntesis For RT-PCR SSâ„¢ II RTâ€, de InvitrogenTM. Se usaron primers específicos para dos genes endógenos: cox y actina. El primero es un gen mitocondrial y el segundo es un gen nuclear. Se observaron señales claras y diferenciables de amplificación para cox, utilizando el kit “ONE Step superScriptTMâ€, con un tamaño esperado de 96 pb. Para actina, se observó una señal clara de amplificación de 300 pb, con el kit “SuperScriptâ„¢ First Strand Syntesis For RT-PCR SSâ„¢ II RTâ€.
Palabras clave: Solanum tuberosum; cox, actina; RT-PCR; papa; Solanum tuberosum; cox; actin; RT-PCR; potato.
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Copyright (c) 2008 Revista Colombiana de Biotecnología
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