几种提取转基因木瓜DNA的方法比较

Comparison of several methods for extracting DNA from transgenic papaya

中文关键词:  木瓜；CTAB法；SDS法；试剂盒法；DNA提取  荧光PCR

英文关键词:papaya  CTAB method  SDS method  kit method  DNA extraction  real-time PCR

中文摘要:

      目的 比较CTAB法、SDS法及试剂盒法分别提取木瓜果皮、果肉、种籽DNA的效果, 以提高转基因木瓜的检测准确率。方法 对转基因木瓜内源基因Papain及外源基因NPTII、CP进行普通PCR, 同时对基因表达零件CaMV35S启动子和NOS终止子进行荧光PCR, 以检测提取DNA的质量。结果 分析电泳产物发现, SDS法提取木瓜种籽样品的DNA未能适合普通PCR检测的要求, 其他方法均能满足普通PCR要求。荧光PCR结果显示, 3种方法提取的DNA都满足荧光扩增要求。结论 比较3种提取方法及3种木瓜材料, 发现用CTAB法或SDS法以果皮为材料提取DNA质量最佳。

英文摘要:

      Objective Comparison of several methods for extracting DNA from pericarp, pulp and seed of papaya using CTAB method, SDS method and kit method, so as to improve the detection accuracy of transgenic papaya. Methods Papaya species-specific Papain gene and exogenous gene of NPTII and CP were detected by common PCR, meanwhile CaMV35S promoter and NOS terminator were detected by fluorescent PCR, in order to detect the quality of extracted DNA. Results By analyzing the electrophoresis products, extraction of papaya seeds’ DNA by SDS method was failed to fit the requirements of common PCR detection except other else. The results of fluorescent PCR detection demonstrated that DNA from 3 kinds of papaya materials extracted by 3 kinds of methods all had typical fluorescence amplification curves. Conclusion Compared 3 kinds of extraction methods and 3 kinds of papaya material, CTAB method or SDS method and extracting pericarp DNA had the best quality of extracted DNA.