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- Title
Estandarización de un protocolo sencillo para la extracción de adn a partir de quesos frescos, semi-curados y curados de cabra.
- Authors
Canales, A. M.; Baquero, M. R.; Delgado, J. V.; Martínez, A.
- Abstract
Cheese is the fresh or matured product, solid or semi-solid, obtained from totally or partially skimmed milk, coagulated totally or partially by the action of rennet, by an acidification process or by the addition of a proteolytic enzyme (enzymatic coagulation). Many goat cheeses are made with milk from a specific breed and this is stated in their specifications as part of the protected designations of origin, however, there is no traceability system for this factor. Genetic traceability is based on the identification of animals and their products through the study of specific DNA sequences that vary between individuals and allow them to be distinguished from one another. The future of native breeds depends on the recognition and valuation of their products. The availability of a method for identifying the breed that produces the milk used in the production of cheese would be an important tool for preventing fraud in these protected products linked to a specific genotype. The availability of a DNA extraction technique with which a sufficient concentration and purity can be obtained at a low cost, without much work effort in the laboratory and in a short time is of interest to many researchers and technicians. The objective of the present study was the standardization of a DNA extraction method, which would give a good quality from different ripening of goat cheeses. Fifteen samples of 100% goat milk cheese with different types of ripening were selected: 5 fresh cheeses, 5 semi-cured cheeses and 5 cured cheeses. In the extraction protocol, an Edwards incubation buffer (Tris 10mM, pH = 8; EDTA20mM, 0.5% SDS, NaCl 2mM), 2-Mercaptoethanol-d6, a modification of Guanidine Thiocyanate lysis buffer (NH2C(=NH)NH2 - HSCN) and silica column are used. After the extraction process was completed, quantification was performed using 2 µL of extracted DNA, obtaining DNA concentration and absorbance ratios with the parameters of 260/230 and 260/280. With the modified and standardized DNA extraction method, sufficient DNA concentrations were obtained, ranging from 21.05 to 214.70 ng/µL. A simple and inexpensive protocol has been standardized for the extraction of genomic DNA from cheese samples, with any type of maturity and which allows obtaining no impurities and high concentrations of DNA for genomic analysis.
- Subjects
NUCLEIC acid isolation methods; GOAT cheese; DNA analysis; IDENTIFICATION of animals; PROTEOLYTIC enzymes; MILK proteins
- Publication
Archivos de Zootecnia, 2022, Vol 71, Issue 275, p200
- ISSN
0004-0592
- Publication type
Article