Assessment of DNA extraction methodologies from three types of equine biological simples
The objective of the research was to analyze four methods of extraction of genomic DNA (gDNA) using a commercial extraction kit, isoamyl phenol-chloroform, Chelex®, and Proteinase K, using samples of whole blood, blood-soaked in filter paper and hair from horses, obtaining a total of sixty samples. Quality and quantity of DNA were determined through spectrophotometry. After verifying that the distribution of the observations did not fit the normal with the Kolmogorov - Smirnov test, the data were analyzed by non-parametric statistics using inferential analysis using Kruskal Wallis. The results demonstrated variations in concentrations and purities; the highest concentration was obtained with isoamyl phenol-chloroform from whole blood (120.36 ng/µL). Using whole blood and filter paper, the means were lower, both for proteinase K (15.71 ng/µL) and for isoamyl phenol-chloroform (29.90 ng/µL). Regarding purity, the Chelex®, isoamyl phenol-chloroform, and proteinase K methods presented higher contaminations, while with the Extraction Kit, the purity values were optimal for the three types of samples. Genomic DNA extraction was achieved using the four protocols, although its validation with a greater number and diversity of samples is necessary.
| Main Authors: | , , , , , , |
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| Format: | Digital revista |
| Language: | spa |
| Published: |
Dirección de Investigación Científica y Tecnológica
2023
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| Online Access: | https://revistascientificas.una.py/index.php/comp/article/view/3882 |
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