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Development of a duplex qPCR assay with locked nucleic acid probes for A, B and E kappa-casein variants detection
dc.creator | Jiménez Montenegro, Lucía | es_ES |
dc.creator | Mendizábal Aizpuru, José Antonio | es_ES |
dc.creator | Alfonso Ruiz, Leopoldo | es_ES |
dc.creator | Azparren Domínguez, Leire | es_ES |
dc.creator | Urrutia Vera, Olaia | es_ES |
dc.date.accessioned | 2023-04-20T17:59:59Z | |
dc.date.available | 2023-04-20T17:59:59Z | |
dc.date.issued | 2022 | |
dc.identifier.citation | Jiménez-Montenegro, L., Mendizabal, J. A., Alfonso, L., Azparren, L., & Urrutia, O. (2022). Development of a duplex qPCR assay with locked nucleic acid probes for A, B and E kappa-casein variants detection. Scientific Reports, 12(1), 16387. https://doi.org/10.1038/s41598-022-20586-w | en |
dc.identifier.issn | 2045-2322 | |
dc.identifier.uri | https://hdl.handle.net/2454/45146 | |
dc.description.abstract | Milk proteins determine important milk technological characteristics. Among caseins, Ƙ-casein has been correlated with fat and protein content and cheese yield. Fourteen Ƙ-caseins variants have been described but the alleles A, B and E are the most important ones due to their frequency and/or influence on the technological aptitudes of milk. Therefore, in the present study two different duplex qPCR assays with locked nucleic acid probes (for positions 13104 and 13124 of the Ƙ-casein gene) were developed for the detection of A, B and E variants. Firstly, DNA isolation method from milk somatic cells and hair was optimised. The developed 13124-qPCR assay showed an increased sensitivity reaching up to 6.7 copies DNA copies/reaction at a 95% confidence level with A, B and E alleles reference samples. The 13104-qPCR assay reached up to 6.7 DNA copies/reaction for A allele reference sample and 67 DNA copies/reaction for B and E samples. Intra-assay variation results were below 6%. Applicability was determined using DNA samples from animals with known genotype for Ƙ-casein (AA, AB, BB, BE, AE, EE) and both assays were able to discriminate among the six genotypes with 100% accuracy. Thus, this qPCR method represents a sensitive and rapid option for the detection of Ƙ-casein alleles in both hair and milk samples. | en |
dc.format.mimetype | application/pdf | en |
dc.language.iso | eng | en |
dc.publisher | Springer Nature | en |
dc.relation.ispartof | Scientific Reports (2022) 12, 16387 | en |
dc.rights | © The Author(s) 2022. This article is licensed under a Creative Commons Attribution 4.0 International License. | en |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.subject | Duplex qPCR assay | en |
dc.subject | Locked nucleic acid probes | en |
dc.subject | K-casein | en |
dc.title | Development of a duplex qPCR assay with locked nucleic acid probes for A, B and E kappa-casein variants detection | en |
dc.type | Artículo / Artikulua | es |
dc.type | info:eu-repo/semantics/article | en |
dc.date.updated | 2023-04-20T17:51:33Z | |
dc.contributor.department | Agronomía, Biotecnología y Alimentación | es_ES |
dc.contributor.department | Agronomia, Bioteknologia eta Elikadura | eu |
dc.contributor.department | Institute on Innovation and Sustainable Development in Food Chain - ISFOOD | en |
dc.rights.accessRights | Acceso abierto / Sarbide irekia | es |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | en |
dc.identifier.doi | 10.1038/s41598-022-20586-w | |
dc.relation.publisherversion | https://doi.org/10.1038/s41598-022-20586-w | |
dc.type.version | Versión publicada / Argitaratu den bertsioa | es |
dc.type.version | info:eu-repo/semantics/publishedVersion | en |