Mura, Maria Consuelo and Daga, Cinzia and Bodano, Sara and Paludo, Marta and Luridiana, Sebastiano and Pazzola, Michele and Dettori, Maria Luisa and Vacca, Giuseppe Massimo and Carcangiu, Vincenzo (2013) Development of a RNA extraction method from milk for gene expression study in the mammary gland of sheep. Molecular Biology Reports, Vol. 40 (3), p. 2169-2173. ISSN 0301-4851. eISSN 1573-4978. Article.
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The aim of the study was to develop a reliable method for the RNA extraction from milk of Sarda sheep breed and to highlight if the extracted RNA can be used for expression study on mammary genes involved in milk fat synthesis using RT-qPCR. The main result is that a sample of 150 ml of milk provides an optimal amount of RNA (73.5 μg/ml). The highest RNA concentration has been found in the samples analysed within 4 h after collection. The RNA extracted was positively correlated to the number of somatic cells (P < 0.001). The efficiency of the extraction method was confirmed by the results obtained from qPCR which showed a Ct value, for SREBPF1 gene of 26.8 ± 0.15. This research demonstrated that the high-quality of the RNA obtained is suited to use for studies of mammary genes expression in sheep, avoiding any damage caused by mammary gland biopsy.
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