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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Ca MV35S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。
Objective To compare the effects of CTAB method, SDS method and kit method on the extraction of papaya peel, pulp and seed DNA respectively to improve the detection accuracy of transgenic papaya. Methods The common papaya (Papain) and exogenous gene NPTII (CP) of transgenic papaya were cloned by PCR, and the CaMV35S promoter and NOS terminator of the gene expression part were also detected by fluorescence PCR to detect the quality of extracted DNA. Results Analysis of the electrophoresis products showed that DNA extracted from papaya seed samples by SDS method was not suitable for normal PCR detection. Other methods could meet the requirements of ordinary PCR. Fluorescent PCR results showed that the DNA extracted by the three methods all met the requirements of fluorescence amplification. Conclusion Three extraction methods and three kinds of papaya materials were compared. The results showed that DNA extracted from pericarp by CTAB method or SDS method was the best.