New Rapid Method of DNA Isolation from Milk Somatic Cells

Abstract

Isolation of genomic DNA is one of the basic steps in many different molecular analyses. There are a few reports on methods of DNA isolation from milk, but many of them are time consuming and expensive, and require relatively large volumes of raw milk. In this study a rapid, sensitive, and efficient method of DNA extraction from milk somatic cells of various mammals (cattle, sheep, goats, horses) is presented. It was found that milk is a good source of genomic DNA, and to obtain a sufficient amount and quality of DNA, suitable for molecular analysis such as PCR, 10 mL of raw milk is sufficient. Thanks to this method, stress in animals can be reduced during collection of researched material. Therefore, this method could be widely used in molecular analyses.

Changes in physicochemical properties and DNA quality of milk as affected by different heat treatments

“…For the raw milk, the resulting ratio (1.60) was lower than that (1.79) reported by Pokorska et al . (), but similar to that (1.65) reported by Liu et al . ().…”

“…Theoretically, pure DNA solutions have an A 260 /A 280 of above 1.8, but we found that the ratio of absorbance (A 260 /A 280 ) of all DNA solutions was below 1.8, probably due to contamination by phenols or proteins. For the raw milk, the resulting ratio (1.60) was lower than that (1.79) reported by Pokorska et al (2016), but similar to that (1.65) reported by Liu et al (2014). These differences in purity of the raw milk samples might be attributed to the different extraction method we used.…”

Single nucleotide polymorphisms in the CXCR1 gene and its association with clinical mastitis incidence in Polish Holstein-Friesian cows

“…Genomic DNA was isolated from the cows' milk, using the method developed at the Department of Cattle Breeding, Faculty of Animal Sciences, University of Agriculture in Kraków (Pokorska et al, 2016). The qualitative and quantitative analysis of the isolated DNA was carried out using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, Massachusetts USA).…”

Qualitative and quantitative identification of adulteration of milk powder using DNA extracted with a novel method

“…A successful PCR assay depends largely on the quality of extracted DNA; thus, extraction of high-quality DNA has been a crucial step in the authentication process. Numerous DNA extraction methods have been used for the preparation of DNA from milk, including research protocols and commercial kits (Liu et al, 2014;Volk et al, 2014;Pokorska et al, 2016), but the DNA extraction method for milk powder is still poorly defined. As adulteration of milk powder increases, it is necessary to develop an effective DNA extraction method suitable for PCR-based investigations of milk powder.…”