Mejoras en un método comercial de extracción de ADN para obtener extractos de ácido nucleico de alta calidad a partir de yemas vegetativas de Populus tremuloides Michx.
DOI:
https://doi.org/10.18387/polibotanica.60.12Palabras clave:
populus tremuloides , asilamiento, ADN, amplificacion por PCR, integridad del ADNResumen
RESUMEN
Populus tremuloides es una especie arbórea de interés por el potencial descubrimiento de una historia adaptativa en su genoma, lo que hace esencial aislar correctamente su ADN. Este estudio examinó dos métodos de extracción basados en el kit Nucleo Spin™ Plant II, usando yemas de crecimiento de ocho muestras. El método 1 usó tampón PL1, un homogeneizador y una incubación con RNasa de 30 minutos; el método 2 empleó tampón PL2, perlas de vidrio y una incubación con RNasa de 45 minutos. Después, el ADN fue amplificado por PCR. El método 1 produjo una concentración media de ADN de 137.9 ng/µL y una relación 260/280 de 1.8, pero mostró fragmentación del ADN en gel, lo que puede afectar algunos análisis. El método 2, aunque con menor concentración (65.9 ng/µL) y relación 260/280 de 1.77, presentó una banda clara en el gel, indicando mejor integridad del ADN. La PCR fue exitosa en ambos casos, pero la mayor integridad del ADN en el método 2 sugiere que es más adecuado para estudios que requieren fragmentos intactos. Aunque el método 1 ofrece mayor cantidad y pureza, el método 2 es preferible cuando la integridad del ADN es crítica para el análisis genómico.
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