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[目的]针对现有的泰国茉莉香米品种纯度检测方法的不足,建立特异、灵敏、准确、具有实用性的泰国茉莉香米品种纯度检测方法。[方法]通过对影响RAPD结果的模板DNA、Mg2+离子、随机引物、dNTPs和Taq酶的浓度条件进行优化、以及随机引物的筛选,建立了泰国茉莉香米法定品种KDML105和RD15的双引物协同RAPD检测方法。[结果]得到最适宜的RAPD反应体系为:反应体积25μl,模板DNA浓度4-32ng/μl、随机引物浓度200μg/L、Mg2+浓度2.0mmo1/L、dNTPs浓度200μmol/L和Taq酶1.0U。通过两条DNA分子标记的双隐性特征可以区分茉莉香米和非茉莉香米品种。[结论]该技术可以有效地弥补了感官法和水煮法的不足,具有操作简单、快速灵敏、费用低等优点,适合在日常检验中推广使用。
[Objective] The aim of the study was to establish a method for purity determination of Thai jasmine rice with specificity, sensitivity, accuracy and practicability according to the deficiency of the existing method for purity detection of Thai jasmine rice. [Method] The double primer of KDML105 and RD15, a common Thai jasmine rice, was established by the combination of RAPD template DNA, Mg2 + ions, random primers, dNTPs and Taq polymerase concentration conditions and random primers screening. Detection method. [Result] The optimal reaction system was as follows: reaction volume 25μl, template DNA concentration 4-32ng / μl, random primer concentration 200μg / L, Mg2 + concentration 2.0mmol / L, dNTPs concentration 200μmol / L and Taq enzyme 1.0U. The two recessive traits of two DNA molecular markers distinguish jasmine rice from non-jasmine rice. [Conclusion] The technique can effectively make up for the lack of sensory and boiling methods, has the advantages of simple operation, quick and sensitive, low cost and is suitable for popularization and use in routine tests.