REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA
In order to compare five volumes of commercial reagent applied in the extraction of bovine DNA in plucked hair, random samples of hair from the tail area of 25 Creole cattle breeds Guaymi and Guabala were taken. DNA concentration in nanograms per microliter (ng/ul) was obtained and the purity was determined by the relationship between absorbance at 260 nm and 280 nm (R) for each of the samples. The results of the analysis conducted for treatments showed no significant differences for the value of R. However, for concentration, a significant differences between treatment 5 and 4 (P = 0.019) and treatment 5 and 1 (P = 0.021) were observed. The overall average of R was 1,77 ± 0,22 and concentration was 34,96 ± 7,18 ng/ul. The observed values of R ranging from 1,37 ± 0,60 in treatment 3 and 2,24 ± 0,61 in treatment 2. As to the DNA concentration, the values were in 12,28 ± 12,98 ng/ul in the treatment 4 and 75,21 ± 14,04 ng/ul in the treatment 5. Genomic DNA extraction was achieved in all treatments with similar values of purity. Optimization occurred up to five times extraction capacity of the commercial product and reduced cost per sample; also the sequence of steps of the protocol using a single thermal cycler in the total extraction process was simplified.
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Instituto de Innovación Agropecuaria de Panamá
2016
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oai:ojs.revistacienciaagropecuaria.ac.pa:article-1002021-07-06T16:21:03Z REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA RENDIMIENTO DE PROTOCOLO DE EXTRACCIÓN DE ADN DE BOVINOS CRIOLLOS Villalobos–Cortés, Axel González–Herrera, Rita In order to compare five volumes of commercial reagent applied in the extraction of bovine DNA in plucked hair, random samples of hair from the tail area of 25 Creole cattle breeds Guaymi and Guabala were taken. DNA concentration in nanograms per microliter (ng/ul) was obtained and the purity was determined by the relationship between absorbance at 260 nm and 280 nm (R) for each of the samples. The results of the analysis conducted for treatments showed no significant differences for the value of R. However, for concentration, a significant differences between treatment 5 and 4 (P = 0.019) and treatment 5 and 1 (P = 0.021) were observed. The overall average of R was 1,77 ± 0,22 and concentration was 34,96 ± 7,18 ng/ul. The observed values of R ranging from 1,37 ± 0,60 in treatment 3 and 2,24 ± 0,61 in treatment 2. As to the DNA concentration, the values were in 12,28 ± 12,98 ng/ul in the treatment 4 and 75,21 ± 14,04 ng/ul in the treatment 5. Genomic DNA extraction was achieved in all treatments with similar values of purity. Optimization occurred up to five times extraction capacity of the commercial product and reduced cost per sample; also the sequence of steps of the protocol using a single thermal cycler in the total extraction process was simplified. Con el objetivo de comparar cinco volúmenes de reactivo comercial aplicado en la extracción de ADN en pelo de bovino, se tomaron muestras aleatorias de pelo de la zona caudal de 25 bovinos criollos de la razas Guaymí y Guabalá. Se obtuvo la concentración de ADN en nanogramos por microlitro (ng/ul) y se determinó la pureza mediante la relación entre la absorbancia 260 nm y 280 nm (R) para cada una de las muestras. Los resultados del análisis para los tratamientos realizados, no mostraron diferencias significativas para el valor de R. Sin embargo para la concentración, se observaron diferencias entre el tratamiento 5 y 4 (P=0,019) y tratamiento 5 y 1 (P=0,021). La media general de R fue 1,77 ± 0,22 y para concentración fue de 34,96 ± 7,18 ng/ul. Los valores observados en R oscilaron entre 1,37 ± 0,60 en el tratamiento 3 y 2,24 ± 0,61 en el tratamiento 2. En cuanto a la concentración de ADN, los valores oscilaron en 12,28 ± 12,98 ng/ul en el tratamiento 4 y 75,21 ± 14,04 ng/ul en el tratamiento 5. Se logró extracción de ADN genómico en todos los tratamientos con similares valores de pureza. Se produjo una optimización de hasta cinco veces la capacidad de extracción del producto comercial y una reducción del costo por muestra; además se simplificó la secuencia de pasos del protocolo al utilizar un termociclador en el proceso total de extracción. Instituto de Innovación Agropecuaria de Panamá 2016-11-02 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf http://www.revistacienciaagropecuaria.ac.pa/index.php/ciencia-agropecuaria/article/view/100 Ciencia Agropecuaria; Núm. 25 (2016); 108–117 2414-3278 0258-6452 spa http://www.revistacienciaagropecuaria.ac.pa/index.php/ciencia-agropecuaria/article/view/100/70 |
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| author |
Villalobos–Cortés, Axel González–Herrera, Rita |
| spellingShingle |
Villalobos–Cortés, Axel González–Herrera, Rita REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA |
| author_facet |
Villalobos–Cortés, Axel González–Herrera, Rita |
| author_sort |
Villalobos–Cortés, Axel |
| title |
REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA |
| title_short |
REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA |
| title_full |
REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA |
| title_fullStr |
REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA |
| title_full_unstemmed |
REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA |
| title_sort |
removal performance protocol creole cattle dna |
| description |
In order to compare five volumes of commercial reagent applied in the extraction of bovine DNA in plucked hair, random samples of hair from the tail area of 25 Creole cattle breeds Guaymi and Guabala were taken. DNA concentration in nanograms per microliter (ng/ul) was obtained and the purity was determined by the relationship between absorbance at 260 nm and 280 nm (R) for each of the samples. The results of the analysis conducted for treatments showed no significant differences for the value of R. However, for concentration, a significant differences between treatment 5 and 4 (P = 0.019) and treatment 5 and 1 (P = 0.021) were observed. The overall average of R was 1,77 ± 0,22 and concentration was 34,96 ± 7,18 ng/ul. The observed values of R ranging from 1,37 ± 0,60 in treatment 3 and 2,24 ± 0,61 in treatment 2. As to the DNA concentration, the values were in 12,28 ± 12,98 ng/ul in the treatment 4 and 75,21 ± 14,04 ng/ul in the treatment 5. Genomic DNA extraction was achieved in all treatments with similar values of purity. Optimization occurred up to five times extraction capacity of the commercial product and reduced cost per sample; also the sequence of steps of the protocol using a single thermal cycler in the total extraction process was simplified. |
| publisher |
Instituto de Innovación Agropecuaria de Panamá |
| publishDate |
2016 |
| url |
http://www.revistacienciaagropecuaria.ac.pa/index.php/ciencia-agropecuaria/article/view/100 |
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