快速提取用于PCR分析的几种蔬菜DNA的方法  

RAPID EXTRACTION METHODS OF VEGETABLE DNA SUITABLE FOR PCR ANALYSIS

作　　者：孟祥栋[1] 马红[1] 盖树鹏[1] 

机构地区：[1]山东农业大学园艺系,山东泰安271018

出　　处：《应用与环境生物学报》1998年第3期251-254,共4页Chinese Journal of Applied and Environmental Biology

基　　金：国家教委留学回国人员启动基金;山东农业大学科学基金

摘　　要：利用两种不同的简便方法提取大白菜、大葱和厚皮酣瓜发芽种子的DNA，均能产生具有重现性的RAPD结果这两种方法提取DNA的PCR扩增结果与多次利用苯酚／氯仿提取DNA的结果一致.RNase处理与否对提取DNA的PCR扩增结果没有影响.方法1不用氯仿，但残留蛋白质等容易造成DNA溶解困难；方法2提取的DNA纯度高对于含蛋白质较少的植物器官或组织，可以选用方法1，而对于含蛋白质较多的植物器官或组织，宜选用方法2.两种方法提取的DNA量与用苯酚／氯仿提取DNA的量相似样品之间提取DNA量的多少主要受研磨破碎程度高低所决定.方法1每人每天提取约120个样品，方法2提取约100个样品，比先前的方法快1倍多.所提取的每个样品DNA约能用于100次PCR反应，两种DNA提取方法简便、迅速，为快速进行大量样品的PCR分析提供了条件.Two different simple methods were tised to extract DNA from the germinated seeds of Chinese cabbage, Welsh onion and melon. Reproducible and consistent RAPD results were achieved with DNA isolated by the two methods. The amplification results were the same as those with DNA from phenol/chloroform extraction methods. RAPD results were not affected by RNase beatment of the DNA compared with those without treatment. Method 1 was simpler than method 2 and chloroform was not used in method 1. Protein and other residues were easily mixed with DNA using method 1, but it was more difficult to dissolve DNA. Method 2 could obtain cleaner DNA. Method 1 was more suitable for plant materials containing less protein and method 2 was more suitable for the materials containing more protein. DNA quantity extracted by the two methods was similar to that by phenol/chloroform methods. The main cause for quantity differences was the ground levels of samples. The finer the sample was ground, the more the DNA was isolated. About 120 samples could be finished by one person with method 1 and about 100 with method 2. DNA extracted by the two methods could be used for abotlt 100PCR reactions. The two methods were simple. fast and reproducible, and made fast PCR analyses of many samples possible.

关 键 词：DNA提取 PCR 蔬菜 

分 类 号：Q781[生物学—分子生物学]