一种适于PCR扩增的茶树基因组DNA快速提取方法

A Rapid Extraction Method of Tea Plant Genomic DNA Suitable for PCR Amplification

为了建立一种从茶树嫩叶中快速提取基因组DNA的方法,笔者通过采用常温研磨、常温裂解、一步抽提等措施对CTAB提取法进行了改良和简化,省去了DNA纯化和使用RNase酶降解RNA两个环节。使用该方法以茶树嫩叶为材料提取基因组DNA,并进行紫外光谱法、琼脂糖电泳、PCR扩增等方式检测。结果表明,该方法提取的DNA,其260 nm/280 nm吸光度比值介于1.81~1.84,260 nm/230 nm吸光度在1.75~1.88;经琼脂糖凝胶电泳检测,获得的DNA条带较亮且无明显拖尾现象;利用ISSR引物进行PCR检测,能够获得清晰稳定条带。说明该方法操作快速简便,提取的基因组DNA能够满足PCR反应需要。

In order to establish a method for rapid extraction of genomic DNA from tea leaves,the CTAB extraction method was improved and simplified by grinding at room temperature,splitting at room temperature and one-step extraction,eliminating two steps of DNA purification and RNA degradation by RNase.Using this method,the genomic DNA was extracted from the tender leaves of tea plant,and detected by UV,agarose electrophoresis and PCR.The results showed that the genomic DNA extracted from this method,its absorbance ratio of 260 nm/280 nm was between 1.81 and 1.84,and that of 260 nm/230 nm was between 1.75 and 1.88.The DNA bands obtained by agarose gel electrophoresis were brighter and had no obvious tail phenomenon.Using ISSR primers for PCR detection,a clear and stable DNA band could be obtained.The results show that the method is rapid and simple,and the extracted genomic DNA can meet the needs of PCR reaction.