[Objective] The aim was to establish more specific, sensitive, accurate and practical method to detect purity of Thai Hom Mali rice. [Method] RAPD method cooperated with two primers of KDML105 and RD15 was established...[Objective] The aim was to establish more specific, sensitive, accurate and practical method to detect purity of Thai Hom Mali rice. [Method] RAPD method cooperated with two primers of KDML105 and RD15 was established by feeling for the extraction methods of DNA, optimizing concentrations of the factors influencing the results of RAPD such as template DNA, Mg2+, random primer, dNTPs and Taq polymerase, and screening the random primers. [Result] The optimum RAPD reaction system was 25.0 μl in total volume, containing 4.0-32.0 ng/μl of template DNA, 200.0 μg/L random primer, 2.0 mmol/L Mg2+, 200.0 μmol/L dNTPs and 1.0 U of Taq enzyme. Then, the Thai Hom Mali rice and non-Thai Hom Mali rice can be distinguished according to the presence or absence of two DNA markers. [Conclusion] The RAPD technology can effectively cover the shortages of identifications by sense and boiling in water; in addition, it is simple, sensitive and low-cost, suitable to be used in routine tests.展开更多
文摘[Objective] The aim was to establish more specific, sensitive, accurate and practical method to detect purity of Thai Hom Mali rice. [Method] RAPD method cooperated with two primers of KDML105 and RD15 was established by feeling for the extraction methods of DNA, optimizing concentrations of the factors influencing the results of RAPD such as template DNA, Mg2+, random primer, dNTPs and Taq polymerase, and screening the random primers. [Result] The optimum RAPD reaction system was 25.0 μl in total volume, containing 4.0-32.0 ng/μl of template DNA, 200.0 μg/L random primer, 2.0 mmol/L Mg2+, 200.0 μmol/L dNTPs and 1.0 U of Taq enzyme. Then, the Thai Hom Mali rice and non-Thai Hom Mali rice can be distinguished according to the presence or absence of two DNA markers. [Conclusion] The RAPD technology can effectively cover the shortages of identifications by sense and boiling in water; in addition, it is simple, sensitive and low-cost, suitable to be used in routine tests.