Valutazione della possibilità di utilizzo del DNA estratto da campioni di latte dei controlli funzionali per l'attribuzione della paternità tramite Microsatelliti nei piccoli ruminanti

The aims of the present work were to get ready an economic, rapid and toxic solvent-free method to extract DNA from sheep milk somatic cells and to set up a system of microsatellites for paternity assignment in Sarda sheep breed. During the years, several techniques which describe the isolation of genomic DNA from somatic cells of cow and goat milk have been reported. Milk is preferred to blood as a source of DNA because its collection is routinely performed, less expensive and more easily accomplished than blood collection. Moreover milk use limits stressful practices such as capture, handling and venipuncture in animal management. A rapid alkaline method resulted most suitable as method for DNA extraction from sheep milk because fast and economic.Genomic DNA was extracted from somatic cells starting from 20 mL of sheep milk and used as substrate for the polymerase chain reaction. A system of 10 highly informative microsatellites (MCM058, LSCV06, BM6444, BMS2213, CSSM43, BMS2252, MCM120, OLADRB, MCM373, BMS0360) was chosen and spread in 3 multiplex for paternity assignment in the Sarda breed. The proposed system would be a useful alternative to traditional pedigree recording in the Sarda breed. Not only it had a high probability of excluding incorrect parental (Pe=0.99996) relationships but it also provided a high percentage of unequivocal paternity assignment (P=1.0 in a simulated population). Its regular utilization would increase the number of individuals with known ancestors in the Sarda breed, with favorable consequences on the number of genetically evaluated animals and the precision of their breeding values estimation. OLADRB, MCM373, BMS0360) was chosen and spread in 3 multiplex for paternity assignment in the Sarda breed. The proposed system would be a useful alternative to traditional pedigree recording in the Sarda breed. Not only it had a high probability of excluding incorrect parental (Pe=0.99996) relationships but it also provided a high percentage of unequivocal paternity assignment (P=1.0 in a simulated population). Its regular utilization would increase the number of individuals with known ancestors in the Sarda breed, with favorable consequences on the number of genetically evaluated animals and the precision of their breeding values estimation.