野油菜黄单胞菌基因组DNA快速提取方法和酶切

The Rapid Extracting Approach and Digestion of the Genomic DNA from Xanthomonas campestris

本文建立了一种野油菜黄单胞菌(Xanthomonas campestris)基因组DNA的简便快速提取方法。用去污剂SDS和CTAB破坏细胞膜结构,使胞内核酸释放,用酚/氯仿/异戊醇去除蛋白质,经异丙醇沉淀DNA,TE溶解DNA。采用此SDS和CTAB结合的方法提取的DNA琼脂糖电泳图谱与试剂盒UNIQ-10提取的基因组DNA及D-5010A试剂盒法提取的基因组DNA图谱相同,DNA经紫外分光光度计检测OD260/OD280在1.80～1.90之间。DNA能被EcoRI、XbalI酶切消化。本方法能快速简便地提取野油菜黄单胞菌DNA,提取的DNA含量较高、纯度较好,比试剂盒更经济,也可用于酶切分析、构建文库和PCR扩增。

A rapid approach in the genomic DNA extracting from the Xanthomonas campestris was promoted in this paper. Thedetergent SDS and CTAB were used to destroy the structure of cell membrance and the cellular nucleic acids was liberated, andthen, the proteins; were removed by using pheno, chloroform, isoamyl alcohol solution, the DNA was precipitated and dissolvedby using isoproanol and TE buffer respectively. Results showed that the DNA map of agarose electrophoresis was the same asthose generated from the UNIQ-10 and D-5010A approaches. The OD260/OD280 was in 1.80～1.90.The DNA was able to bedigested by the EcoRI, XbalI. This approach could used in the genomic DNA extracting from Xanthomonas campestris rapidly.The purity of the obtained DNA was good enough for the further digestion by restriction edonucleases, the establishment ofgenomic library and the PCR analysis.