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以巨峰葡萄叶片为试材 ,在CTAB法、SDS法及高盐低pH法基础上加以改进 ,应用 5种方法对葡萄DNA提取、纯化方法进行了比较研究。结果表明 ,高盐低pH法和区室化提取法是 2种适合于葡萄属植物DNA提取的方法。这 2种方法提取的DNA纯度高 ,凝胶电泳显示无明显降解现象 ,能被限制性内切酶酶切 ,适宜作为PCR模板应用 ,并成功地进行了扩增。还对影响葡萄等富含多酚、多糖等物质的多年生果树DNA提纯的有关因素、β Me及PVP的抗氧化作用等问题进行了探讨
Taking Kyoho grapevine leaves as test material, the methods of CTAB, SDS and high salt and low pH were improved. Five methods were used to compare the DNA extraction and purification of grape. The results showed that high salt and low pH and compartmentalized extraction were two suitable methods for DNA extraction from Vitis. The DNA extracted by these two methods has high purity and no obvious degradation phenomenon by gel electrophoresis. It can be digested by restriction endonuclease and suitable for PCR template application and has been successfully amplified. The factors affecting DNA purification of perennial fruit trees such as grape and other polyphenols and polysaccharides, antioxidation effects of β Me and PVP were also discussed