5种提取方法对甘蔗汁中DNA提取的效果
|
摘要
[目的]研究5种提取方法对甘蔗汁中DNA提取的效果。[方法]以甘蔗种茎中获得的蔗汁为材料,采用2种植物基因组提取试剂盒柱式法和磁珠法、SDS法、CTAB法及简化提取法5种方法提取DNA,对比所提取的DNA浓度、纯度及后续PCR扩增的影响。[结果]以SDS法提取获得的蔗汁DNA浓度最高,平均可达377.50 ng/m L。而柱式法的试剂盒,提取的DNA浓度最低,PCR检测没有能够获得特异的目标条带,不适合用于蔗汁DNA的直接提取。虽然简化法提取的DNA浓度较低,但是能够满足PCR扩增的要求,由于不涉及氯仿等有害物质,操作简单,适用于对DNA质量要求不高时的快速提取。[结论]SDS法最适合用于蔗汁中DNA的提取。
[Objective]To study the effects of five methods on total DNA extraction from sugarcane juice. [Method]In this study,five methods,including two kinds of the plant DNA extraction kit( with column and magnetic bead,respectively),CTAB method,SDS method and simplified DNA extraction method were used to extract DNA from sugarcane juice collected from seed-stem,and the quality and quantity of DNA and the following PCR analysis were compared. [Result]SDS method was appropriate for DNA extraction from sugarcane juice and obtained highest amount of DNA from sugarcane juice( 377. 50 ng/μL,on average). DNA yield from the plant DNA extraction kit( with column) was lowest and might interfere the following PCR analysis,so this method was not suitable for directly extracting DNA from sugarcane juice. The simplified DNA extraction method had advantage of simple and fast. Although its yield and purity were lower than those extracted by SDS and CTAB methods,it meet the demand of the following PCR analysis and was suitable for fast extraction with lower quality requirement.[Conclusion]SDS method was appropriate for DNA extraction from sugarcane juice.
[Objective]To study the effects of five methods on total DNA extraction from sugarcane juice. [Method]In this study,five methods,including two kinds of the plant DNA extraction kit( with column and magnetic bead,respectively),CTAB method,SDS method and simplified DNA extraction method were used to extract DNA from sugarcane juice collected from seed-stem,and the quality and quantity of DNA and the following PCR analysis were compared. [Result]SDS method was appropriate for DNA extraction from sugarcane juice and obtained highest amount of DNA from sugarcane juice( 377. 50 ng/μL,on average). DNA yield from the plant DNA extraction kit( with column) was lowest and might interfere the following PCR analysis,so this method was not suitable for directly extracting DNA from sugarcane juice. The simplified DNA extraction method had advantage of simple and fast. Although its yield and purity were lower than those extracted by SDS and CTAB methods,it meet the demand of the following PCR analysis and was suitable for fast extraction with lower quality requirement.[Conclusion]SDS method was appropriate for DNA extraction from sugarcane juice.
引文
[1]杨海霞,吴弦华,农秋连,等.甘蔗分子标记应用研究进展[J].中国糖料,2016,38(6):57-61.
[2]仓晓燕,黄应昆,李文凤,等.甘蔗抗褐锈病基因与分子标记研究进展[J].中国糖料,2016,38(6):62-65.
[3]刘昔辉,宋焕忠,张革民,等.一种快速高效提取甘蔗及其近缘属基因组DNA的方法[J].西南农业学报,2010,23(2):515-518.
[4]田春艳,王先宏,李富生,等.割手密线粒体DNA的简易快速提取方法[J].分子植物育种,2015,13(5):1141-1145.
[5]黄东亮,覃肖良,廖青,等.高质量甘蔗基因组DNA的简便快速提取方法研究[J].生物技术通报,2010(5):101-106.
[6]王英,邱海燕,高和琼,等.甘蔗基因组DNA提取方法的研究[J].中国农学通报,2008,24(12):44-49.
[7]李建勇,蒙姣荣,陈保善.从甘蔗种茎茎汁中提取基因组DNA的方法[J].中国农学通报,2010,26(14):86-89.
[8]XIN Z G,VELTEN J P,OLIVER M J,et al.High-throughput DNA extraction method suitable for PCR[J].Biotechniques,2003,34(4):820-824,826.
[9]肖乃衍,林冰,刘迪,等.甘蔗内标基因PCR检测技术研究[J].热带作物学报,2017(3):500-507.
[10]LING H,WU Q B,GUO J L,et al.Comprehensive selection of reference genes for gene expression normalization in sugarcane by real time quantitative RT-PCR[J].PLo S One,2014,9(5):97469.
[11]张娟,吴炜亮,谭嘉力,等.植物源性转基因食品DNA提取技术的研究进展[J].食品工业科技,2015,36(7):396-400.
[12]裴杰萍,端青.DNA提取方法的研究进展[J].微生物学免疫学进展,2004,32(3):76-78.
[13]赵雅楠,王颖,张东杰,等.食品中植物基因组DNA提取方法的分类及其研究进展[J].现代农业科技,2015(22):275-277.
[14]TAYLOR J I,HURST C D,DAVIES M J,et al.Application of magnetite and silica-magnetite composites to the isolation of genomic DNA[J].J Chromatogr A,2000,890(1):159-166.
[15]马晓凡,李林.金针菇DNA提取方法比较[J].安徽农业科学,2017,45(2):158-160.
[16]齐玲倩,刘秀,丁梦璇,等.果汁DNA提取方法的比较[J].食品与发酵工业,2015,41(12):224-228.
[17]任君安,王国兰,程曦,等.果蔬汁饮料DNA提取方法的比较研究[J].食品科技,2013,38(2):42-45.
[18]隋志伟,余笑波,王晶,等.转基因水稻种子核酸提取方法的比较研究[J].中国稻米,2011,17(6):40-43.
[19]姚伟,耿广良,余爱丽,等.一种改良的转基因甘蔗基因组DNA提取方法[J].热带亚热带植物学报,2004,12(3):257-260.
[2]仓晓燕,黄应昆,李文凤,等.甘蔗抗褐锈病基因与分子标记研究进展[J].中国糖料,2016,38(6):62-65.
[3]刘昔辉,宋焕忠,张革民,等.一种快速高效提取甘蔗及其近缘属基因组DNA的方法[J].西南农业学报,2010,23(2):515-518.
[4]田春艳,王先宏,李富生,等.割手密线粒体DNA的简易快速提取方法[J].分子植物育种,2015,13(5):1141-1145.
[5]黄东亮,覃肖良,廖青,等.高质量甘蔗基因组DNA的简便快速提取方法研究[J].生物技术通报,2010(5):101-106.
[6]王英,邱海燕,高和琼,等.甘蔗基因组DNA提取方法的研究[J].中国农学通报,2008,24(12):44-49.
[7]李建勇,蒙姣荣,陈保善.从甘蔗种茎茎汁中提取基因组DNA的方法[J].中国农学通报,2010,26(14):86-89.
[8]XIN Z G,VELTEN J P,OLIVER M J,et al.High-throughput DNA extraction method suitable for PCR[J].Biotechniques,2003,34(4):820-824,826.
[9]肖乃衍,林冰,刘迪,等.甘蔗内标基因PCR检测技术研究[J].热带作物学报,2017(3):500-507.
[10]LING H,WU Q B,GUO J L,et al.Comprehensive selection of reference genes for gene expression normalization in sugarcane by real time quantitative RT-PCR[J].PLo S One,2014,9(5):97469.
[11]张娟,吴炜亮,谭嘉力,等.植物源性转基因食品DNA提取技术的研究进展[J].食品工业科技,2015,36(7):396-400.
[12]裴杰萍,端青.DNA提取方法的研究进展[J].微生物学免疫学进展,2004,32(3):76-78.
[13]赵雅楠,王颖,张东杰,等.食品中植物基因组DNA提取方法的分类及其研究进展[J].现代农业科技,2015(22):275-277.
[14]TAYLOR J I,HURST C D,DAVIES M J,et al.Application of magnetite and silica-magnetite composites to the isolation of genomic DNA[J].J Chromatogr A,2000,890(1):159-166.
[15]马晓凡,李林.金针菇DNA提取方法比较[J].安徽农业科学,2017,45(2):158-160.
[16]齐玲倩,刘秀,丁梦璇,等.果汁DNA提取方法的比较[J].食品与发酵工业,2015,41(12):224-228.
[17]任君安,王国兰,程曦,等.果蔬汁饮料DNA提取方法的比较研究[J].食品科技,2013,38(2):42-45.
[18]隋志伟,余笑波,王晶,等.转基因水稻种子核酸提取方法的比较研究[J].中国稻米,2011,17(6):40-43.
[19]姚伟,耿广良,余爱丽,等.一种改良的转基因甘蔗基因组DNA提取方法[J].热带亚热带植物学报,2004,12(3):257-260.