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牦牛乳中DNA提取方法的建立与优化
引用本文:崔艳华,马莺,曲晓军,李海梅,何胜华.牦牛乳中DNA提取方法的建立与优化[J].哈尔滨工业大学学报,2011,43(2):66-69.
作者姓名:崔艳华  马莺  曲晓军  李海梅  何胜华
作者单位:哈尔滨工业大学食品科学与工程学院;哈尔滨工业大学食品科学与工程学院;黑龙江省科学院微生物研究所;哈尔滨工业大学食品科学与工程学院;哈尔滨工业大学食品科学与工程学院
基金项目:国家自然科学基金资助项目(3087195331074571)
摘    要:为加速牦牛重要基因的挖掘和功能鉴定,以牦牛乳作为试验材料,建立简便的牦牛基因组DNA提取方法.采用常规离心法,从牦牛乳中获得体细胞,进而提取基因组DNA.研究不同去除蛋白方法、不同DNA沉淀剂、不同贮藏条件对牦牛乳中DNA提取质量的影响.DNA质量通过PCR评价.结果表明:高盐法与酚/氯仿法去除蛋白效果相当.高盐法简便...

关 键 词:牦牛乳  体细胞  基因组DNA  提取

Establishment and optimization of the DNA extraction from yak milk
CUI Yan-hu,MA Ying,QU Xiao-jun,LI Hai-mei and HE Sheng-hua.Establishment and optimization of the DNA extraction from yak milk[J].Journal of Harbin Institute of Technology,2011,43(2):66-69.
Authors:CUI Yan-hu  MA Ying  QU Xiao-jun  LI Hai-mei and HE Sheng-hua
Affiliation:School of Food Science and Engineering,Harbin Institute of Technology,150090 Harbin,China;School of Food Science and Engineering,Harbin Institute of Technology,150090 Harbin,China;Institute of Microbiology,Heilongjiang Science Academy,150010 Harbin,China);School of Food Science and Engineering,Harbin Institute of Technology,150090 Harbin,China;School of Food Science and Engineering,Harbin Institute of Technology,150090 Harbin,China
Abstract:A reliable and rapid method for genomic DNA extraction was constructed here to accelerate the identification of important genes in yak.In this method,the yak milk was first centrifuged,and then somatic cells were obtained for DNA extraction.The influences of different methods of protein removal,DNA precipitants,and storage conditions of yak milk on DNA quality were discussed.The quality of DNA was detected using PCR.Experimental results indicated that both the salting-out and phenol-chloroform method were effective for the removal of proteins,however the salting-out method was more simple and rapid.The precipitants isopropyl alcohol and ethanol were similar to precipitate DNA,but the glycol ether was the worst precipitant in this study.The storage conditions of yak milk have remarkable influence on the quality of DNA,and when the yak milk was stored at 4 ℃ for short term,or-20 ℃ for long term,the desired DNA can be got.The DNA extracted from yak milk using the salting-out method can be used in the later genetic research.
Keywords:yak milk  somatic cell  genome DNA  extraction
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