DNA Extraction Methods Used in Leptin’s Identification from Buffolo Milk

Abstract

Leptin is a 16 kDa protein hormone mainly but not exclusively produced and secreted by adipose tissue, which functions as the afferent signal in a feedback loop regulating adipose tissue mass. The progress in animal breeding may be improved by combining traditional performance data with molecular genetic information on quantitative loci in selection index (Buchanan F.C. et al.,2003) The main purpose of this study was to compare different DNA extraction methods needed in PCR. By PCR we can identify leptin’s allele responsible for the quality and production traits of milk. The study material was represented by 30 samples of blood, milk and colostrums taken from buffalos from Salaj and Cluj County. The DNA extraction from blood was done from the leukocytes layer revealed after blood’s coagulation, using the standard protocol written in the paper. The DNA quantity values obtained ranged between 18.9 – 198.4 ng/ µL, and the DNA purity values between 1.07 – 1.76 wavelength 260/280. The DNA extraction from milk’s somatic cells was done using two methods: the rapid method of extraction with PBS, and the classic method. The DNA quantity values obtained through these methods ranged between 14.2 – 156.8 ng/ μL, and the purity values between 1-2 wave lengths 260/280. The DNA quantity and purity values obtained by these three methods were statistically analysed using the ANOVA test and the p values were 0.04476 and 0.02588 which means that there are no significant differences between them. The DNA extraction method from blood can be replaced with succes by these methods of extraction from milk, given the fact that these methods do not imply such a laborious work and uneccessary stress to the buffalos.