Comparison of methods for high quantity and quality genomic DNA extraction from raw cow milk.
Isolation of sufficient quantities of high quality DNA is a prerequisite for molecular studies. Milk somatic cells can be used; however, inhibitors such as fats and proteins make milk a difficult medium for extracting large amounts of quality DNA. We optimized, evaluated and compared three methods, Modified Nucleospin Blood Kit method, Modified TianGen Kit method and Phenol-Chloroform method for genomic DNA extraction from bovine milk. Individual cows' milk and bulk milk samples were collected from a China agricultural university dairy farm. Genomic DNA extracted from each milk sample by the three methods was evaluated for quantity and purity by spectrophotometry and gel electrophoresis, as well as PCR and sequencing. All the three methods were found suitable for genomic DNA isolation from bovine milk, PCR applications, and sequencing. Comparing the three methods, we found that the Modified Nucleospin Blood Kit method was significantly better than the Phenol-Chloroform method in terms of quantity as well as quality (amount, concentration, 260/280 nm and 260/230 nm absorbance ratio), whereas, the Modified TianGen Kit method was more efficient than the Phenol-Chloroform method and cheaper than the Modified Nucleospine Blood Kit method; it yielded reasonably good quantities of good quality DNA and would be suitable for large-scale genotyping of lactating cows.
Key Laboratory of Agricultural Animal Genetics, Breeding and Reprodution, Ministry of Agriculture, National Engineering Laboratory Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, China.
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T
Usman
Key Laboratory of Agricultural Animal Genetics, Breeding and Reprodution, Ministry of Agriculture, National Engineering Laboratory Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, China.
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Y
Yu
Key Laboratory of Agricultural Animal Genetics, Breeding and Reprodution, Ministry of Agriculture, National Engineering Laboratory Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, China.
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C
Liu
Key Laboratory of Agricultural Animal Genetics, Breeding and Reprodution, Ministry of Agriculture, National Engineering Laboratory Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, China.
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Z
Fan
Key Laboratory of Agricultural Animal Genetics, Breeding and Reprodution, Ministry of Agriculture, National Engineering Laboratory Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, China wangyachun@cau.edu.cn.
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Y
Wang
D000818:Animals; D002417:Cattle; D002681:China; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D023281:Genomics / Q000379:methods*; D008892:Milk / Q000737:chemistry*; D016133:Polymerase Chain Reaction
D004247:DNA