Evaluation and comparison of two DNA extraction protocols from three types of sheep samples

Nuñez, Lorena; Rodriguez, María Inés; Gimenez, Guillermo; Martínez, R. D.

doi:10.18004/compend.cienc.vet.2021.11.02.18

Compend. cienc. vet

2021

DOI: 10.18004/compend.cienc.vet.2021.11.02.18

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Evaluation and comparison of two DNA extraction protocols from three types of sheep samples

Lorena Nuñez

María Inés Rodriguez

Guillermo Gimenez

et al.

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2024

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A straightforward technique to obtain genomic DNA from nasal swabs suitable for sheep SNP genotyping analysis

Carracelas,

Peraza,

Ciappesoni

et al. 2024

Agrocienc Urug

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Isolation of high quality and quantity genomic DNA is essential for molecular studies. It is crucial to select a non-invasive and straightforward technique to ensure the efficient collection of DNA, particularly at the farm level. The aim of this study was to determine if nasal swabs are an appropriate biological matrix to obtain good quality genomic DNA suitable for SNP genotyping. In this study, two biological matrices (blood and nasal swabs) were evaluated and compared for the isolation of genomic DNA obtained from 15 female Texel sheep. DNA quality and quantity were assessed using spectrophotometry and gel electrophoreses. Genotype concordance rates were used for comparison. Results showed that the highest concentration mean was obtained from blood samples (159.14 ng/µl), while from nasal swab samples the concentration mean was lower (130.12 ng/µl), but the difference was non-significant. Regarding purity, DNA obtained from nasal swabs presented a higher A260/A280 ratio (1.96), while the one obtained from blood samples was 1.90. Total DNA yield obtained from blood samples (15.91 µg) was significantly higher than the one obtained from nasal swabs (6.51). Blood and nasal swab genotyping concordance rates were high (mean = 0.984). In conclusion, our results indicate that nasal swabs can yield good quality DNA; however, the DNA extraction protocol should be optimized.

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A straightforward technique to obtain genomic DNA from nasal swabs suitable for sheep SNP genotyping analysis

Carracelas,

Peraza,

Ciappesoni

et al. 2024

Agrocienc Urug

View full text Add to dashboard Cite

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Procedimiento para el aislamiento de ADN a partir de sangre coagulada de aves

Arcia,

Marques U,

De La Rosa

et al. 2024

ALPA

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Con la finalidad de aislar ADN genómico de forma económica y segura a partir de muestras de sangre coagulada, se tomaron aproximadamente 250 µL de coagulo de sangre, se agregaron 400 µL de buffer de lisis celular (10 mmol/L Tris-HCl, pH 8,0; 10 mmol/L KCl; 10 mmol/L MgCl2; 2 mmol/L EDTA, pH 8,0; 0,4 mol/L NaCl y 10g/L SDS) y se dejó incubar hasta el día siguiente a temperatura ambiente. Se añadieron 350 µL de cloroformo y se agitó por 10 segundos, después se añadieron 350 µL de acetato de potasio 5M, se agitó por 10 segundos y se centrifugó por 10 minutos (14.000 g). El sobrenadante se transfirió a tubos nuevos y se agregaron 700 µL de isopropanol frio, se centrifugó por 5 minutos a 14.000 g y se descartó el sobrenadante. Posteriormente se realizaron dos lavados con etanol al 70% (700 µL c/u), se centrifugó por 5 minutos y se descartó el líquido. El sedimento se secó 10 min a 45 0C y 8 min a 65 0C para después rehidratar con 20 – 25 µL de TE-1X. La concentración y pureza de los aislados de ADN se determinó mediante espectrofotometría a 260 nm, para un promedio de 2.288,12 ng/µL y 2,04 para la relación A260/A280. La idoneidad del ADN extraído se evaluó mediante la amplificación por PCR de los marcadores MCW0241 y LEI0122. El procedimiento descrito permitió obtener ADN de buena calidad y en grandes cantidades de forma simple, confiable y económica.

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Evaluation and comparison of two DNA extraction protocols from three types of sheep samples

Cited by 2 publications

References 0 publications

A straightforward technique to obtain genomic DNA from nasal swabs suitable for sheep SNP genotyping analysis

A straightforward technique to obtain genomic DNA from nasal swabs suitable for sheep SNP genotyping analysis

Procedimiento para el aislamiento de ADN a partir de sangre coagulada de aves

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