Genomic DNA Extraction and Genotyping

Blood samples were collected from the coccygeal (tail) vein into EDTA Vacutainer tubes (BD Vacutainer System, Becton, Dickinson, and Company, Sparks, MD, USA). Blood samples were centrifuged at 2500×g for 20 min at 4 °C and buffy layers containing white blood cells were collected. Genomic DNA was extracted from the blood buffy coat using the QIAmp DNA Blood Mini Kit according to the manufacturer’s instructions (Qiagen, Hilden, Germany). Purified genomic DNA was quantified spectrophotometrically and subsequently genotyped on the EuroG10K BeadChip at the molecular genetic laboratory service of the Spanish Federation of Holstein Cattle (CONAFE) using the Infinium iScan software for allele assignation (Illumina, San Diego, CA) as previously described^{28 (link)}. The EuroG10K BeadChip is a development by EuroGenomics and its collaborators.

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Canive M., Fernandez-Jimenez N., Casais R., Vázquez P., Lavín J.L., Bilbao J.R., Blanco-Vázquez C., Garrido J.M., Juste R.A, & Alonso-Hearn M. (2021). Identification of loci associated with susceptibility to bovine paratuberculosis and with the dysregulation of the MECOM, eEF1A2, and U1 spliceosomal RNA expression. Scientific Reports, 11, 313.

Publication 2021

BD Vacutainer System QIAmp DNA Blood Mini Kit

Allele Blood Blood buffy coat Cattle Coccygeal Edta Genetic service Genomic Spanish Tail Vein White blood cells

Corresponding Organization :

Other organizations : Euskadiko Parke Teknologikoa, BioCruces Health research Institute, University of the Basque Country, Servicio Regional de Investigación y Desarrollo Agroalimentario

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Variable analysis

independent variables

Sample collection method (coccygeal vein)
Blood sample centrifugation (2500×g for 20 min at 4 °C)
Genomic DNA extraction method (QIAmp DNA Blood Mini Kit)
Genotyping platform (EuroG10K BeadChip)
Allele assignation software (Infinium iScan)

dependent variables

Genomic DNA quantity (measured spectrophotometrically)

control variables

EDTA Vacutainer tubes for blood sample collection
Buffy layer containing white blood cells for DNA extraction

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