醋酸铵法提取卵形鲳鲹基因组DNA

建立了一种简便、安全、经济的提取卵形鲳鲹基因组DNA的方法,即醋酸铵法。该方法采用7.5mol·L~(-1)醋酸铵代替酚、氯仿,通过高盐-SDS去除蛋白和多糖,异丙醇在一定浓度醋酸铵存在条件下选择性沉淀DNA,而不沉淀蛋白,并通过酶切和AFLP验证该法抽提的DNA纯度。该法提取的DNA质量较高,无蛋白质和RNA污染,较酚/氯仿法和试剂盒法抽提得到量多,酶切和AFLP试验结果表明,提取的DNA可用于酶切分析和分子标记等试验。此法安全、简便、经济,该方法更适合于对大量样品的提取。

Ammonium Acetate Method to Isolate High Quality Genomic DNA of Trachinotus ovatus

High-quality DNA was an important base of molecular biology manipulation,such as molecular markers.Ammonium acetate method,a simple,economical and safe method was presented to isolate high quality DNA from Trachinotus ovatus.Instead of phenol and chloroform,7.5 mol·L~（-1） ammonium acetate was used to remove proteins and polysaccharides in an SDS extraction buffer in this method.With a certain concentration of ammonium acetate,isopropyl can selective precipitate DNA without protein.The purity of DNA was validated by restriction enzyme digestion and AFLP analysis.The DNA extraction by ammonium acetate has high-quality, has not protein and RNA pollution,has higher yield compared with phenol/chloroform extraction method and kit method. The results of restriction enzyme digestion and AFLP showed that the purity of the obtained DNA was good enough for the further biotechnology experiment such as digestion by restriction enzyme,molecular marker and so on.This method was safe,simple and economical. This ammonium acetate method was more practical for extraction of a large number of samples.