title: Novel extraction method of genomic DNA suitable for long-fragment amplification from small amounts of milk.
abstract: Isolation of genomic DNA is a prerequisite for assessment of milk quality. As a source of genomic DNA, milk somatic cells from milking ruminants are practical, animal friendly, and cost-effective sources. Extracting DNA from milk can avoid the stress response caused by blood and tissue sampling of cows. In this study, we optimized a novel DNA extraction method for amplifying long (>1,000 bp) DNA fragments and used it to evaluate the isolation of DNA from small amounts of milk. The techniques used for the separation of milk somatic cell were explored and combined with a sodium dodecyl sulfate (SDS)-phenol method for optimizing DNA extraction from milk. Spectrophotometry was used to determine the concentration and purity of the extracted DNA. Gel electrophoresis and DNA amplification technologies were used for to determine DNA size and quality. The DNA of 112 cows was obtained from milk (samples of 13 ± 1 mL) and the corresponding optical density ratios at 260:280 nm were between 1.65 and 1.75. Concentrations were between 12 and 45 μg/μL and DNA size and quality were acceptable. The specific PCR amplification of 1,019- and 729-bp bovine DNA fragments was successfully carried out. This novel method can be used as a practical, fast, and economical mean for long genomic DNA extraction from a small amount of milk.
authors.affiliation: College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi'an, 710062, Shaanxi, China.
authors.forename: Y F
authors.initials: YF
authors.lastname: Liu
authors.affiliation: College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi'an, 710062, Shaanxi, China.
authors.forename: J L
authors.initials: JL
authors.lastname: Gao
authors.affiliation: College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi'an, 710062, Shaanxi, China.
authors.forename: Y F
authors.initials: YF
authors.lastname: Yang
authors.affiliation: College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi'an, 710062, Shaanxi, China.
authors.forename: T
authors.initials: T
authors.lastname: Ku
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: zanlinsen@163.com.
authors.forename: L S
authors.initials: LS
authors.lastname: Zan
mesh_terms: D000818:Animals; D002417:Cattle; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D063427:Food Quality; D023281:Genomics / Q000379:methods*; D008892:Milk / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D017422:Sequence Analysis, DNA
chemical_list: D004247:DNA
keywords: DNA detection; DNA extraction; PCR; milk