RAPD技术在龙须菜遗传多样性研究中的应用 Ⅰ总DNA的提取及RAPD反应条件的优化  

Studies on Application of RAPD to Genetic Diversity in Gracilaria Lemaneiformis Ⅰ. Extraction of DNA and Optimization of RAPD

作　　者：李向峰[1] 隋正红[1] 张学成[1] 

机构地区：[1]青岛海洋大学海洋生命学院

出　　处：《青岛海洋大学学报（自然科学版）》1998年第2期293-298,共6页Journal of Ocean University of Qingdao

基　　金：国家"攀登计划B"项目;曾呈奎海洋科学发展基金

摘　　要：比较两种从龙须菜中提取总ＤＮＡ的方法，这两种方法得到的染色体ＤＮＡ具有较好的完整性，从ＤＮＡ的产量、蛋白质含量等指标可以看出，ＣＴＡＢ法优于蛋白酶Ｋ法。实验对龙须菜ＲＡＰＤ反应条件进行了优化：在２５μＬ反应体系中，含有Ｍｇ２＋２．５ｍｍｏｌ／Ｌ，ｄＮＴＰ１００μｍｏｌ／Ｌ，总ＤＮＡ２５ｎｇ，引物０．２μｍｏｌ／Ｌ和Ｔａｑ酶１Ｕ，经９４℃变性１ｍｉｎ，３５℃退火１ｍｉｎ，７２℃延伸２ｍｉｎ，３５个循环，得到稳定的ＲＡＰＤ扩增图谱。Two methods of DNA extraction from Gracilaria lemaneiformis were compared. The integrity of chromosomal DNA molecules was good for both of the methods. Samples from CTAB method were better than that from proteinase K method in DNA production and protein content. Conditions of RAPD PCR were optimized in the present paper: RAPD reaction in 25μL reaction volumes (containing 2.5m mol/L of Mg 2+ ,100μ mol/L of dNTP, 25ng of total DNAs,0.2μ mol/L of primer and 1 unit of Tag DNA polymerase) was performed smoothly with 35 cycles of denature 1 min at 94℃, annealing 1 min at 35℃ and elongation 2 min at 72℃, and stable RAPD amplification patterns were obtained.

关 键 词：龙须菜 蛋白酶K 脱氧核糖核酸 遗传多样性 

分 类 号：Q949.290.3[生物学—植物学] S644.6[农业科学—蔬菜学]