Novel Extraction Method of Genomic DNA Suitable for Long Fragment Amplification from Small Amounts of Milk
Isolation of amplifiable genomic DNA is a prerequisite for the quality assessment of milk.Milk somatic cells are a practical, animal friendly and cost-effective source of genomic DNA in milking ruminants.Milk as a source to extract DNA, avoiding the use of blood and tissue sampling difficulties do not cause stress response in cows.In this study, a novel DNA extraction method for amplifying long (>1000bp) DNA fragment was optimized and evaluated the isolation of DNA from small amounts of milk samples.We test combined milk somatic cell separation techniques with SDS phenol method for DNA extraction techniques, optimizing and establishing the technology of extracting DNA from milk.Then, spectrophotometer method are used for the determination of DNA concentration and purity, gel electrophoresis and DNA amplification technology are used for DNA size and quality detection.At last, the DNA of 112 cows was obtained from 12mL to 14mL of milk.Their OD260/280 values of purity are among 1.65 to 1.75; concentration are among 121μg/μL to 45μg/μL; sizes and qualities are good; which can be successfully carried out specific PCR amplification of 1019bp and 729bp DNA fragment of bovine.Therefore, this novel method of long genomic DNA extraction from a few of milk is available.It can be used as a practical, fast and economical method of DNA extraction.
milk DNA PCR somatic cells SDS phenol
Yongfeng Liu Junling Gao Yongfang Yang Caixia Ru
College of Food Engineering and Nutritional Science, Shaanxi Normal University, 710062, Xi”an, Shaan Animal Husbandry Company, Xi”an Modem Agriculture Comprehensive Development Corporation, 710018, Xi”
国内会议
第二届中国肉牛选育改良与产业发展国际研讨会暨中国畜牧兽医学会养牛分会八届二次学术研讨会
陕西杨凌
英文
335-340
2013-10-22（万方平台首次上网日期，不代表论文的发表时间）