Modified CTAB Method for Extracting Genomic DNA from Wheat Leaf  

改良CTAB方法快速提取小麦基因组DNA(英文)

作　　者：张晓祥[1] 王玲[1] 寿路路[1] 

机构地区：[1]江苏里下河地区农业科学研究所,江苏扬州225007

出　　处：《Agricultural Science & Technology》2013年第7期946-949,共4页农业科学与技术（英文版）

基　　金：Supported by Major National Transgenic Breeding Project(2011ZX08002-001);the Agricultural Science and Technology Support Program of Jiangsu Province(BE2011306);Agricultural Science and Technology Independent Innovation Fund ofJiangsu Province[CX(12)2026]~~

摘　　要：ObjectiveThe aim was to seek for a rapid DNA minipreparation method from wheat leaf. MethodThe total DNA of wheat leaf was extracted using CTAB, SDS and boiling water, separately, with some modifications. Integrity and purity of nucleic acids were detected through agarose gel electrophoresis, ultraviolet absorption and PCR. ResultThe DNA extracted by the modified CTAB method had high quality and purity, and was not degraded. Two hundreds of DNA samples could be extracted each workday by per capita using this method; and the PCR detection of wheat transgenic plants showed that amplified bands of target gene were clear, without false-positive, and the test results were satisfactory. The DNA purity and concentration extracted by modified SDS method were not as good as that extracted by modified CTAB method, but it also met the DNA requirements of major molecular research. The DNA quantity extracted by modified boiling method was small and there were a lot of impurities in it, PCR detection of this DNA showed no amplified band. ConclusionModified CTAB method is a simple and rapid method for DNA minipreparation from wheat leaf, and was suitable for PCR amplification and other molecular biology researches.[目的]旨在寻求一种微量、快速提取小麦DNA的方法。[方法]以小麦幼叶为材料,分别用改良CTAB法、改良SDS法和沸水浴法提取小麦叶片总DNA,通过琼脂糖凝胶电泳法、紫外吸收法和PCR检测DNA完整性和纯度。[结果]改良CTAB法提取的小麦基因组DNA质量和纯度高、无降解,每人每天可以轻松提取200个DNA样品,对转基因植株的PCR检测显示扩增目标条带清晰一致,无假阳性,试验结果理想。改良SDS法所提DNA纯度和浓度虽略不如改良CTAB法,但仍然符合实验要求。沸水浴法提取量少且杂质多,PCR无有效扩增,结果不理想。[结论]改良CTAB法提供了一种简便、快速微量小麦DNA提取方法,适用于PCR和其他分子生物学研究。

关 键 词：WHEAT DNA extraction Modified CTAB method 

分 类 号：S512.1[农业科学—作物学]