菠菜AFLP反应体系研究(英文)

目的]该研究旨在初步建立适合于菠菜作物的AFLP反应体系。方法]运用改良CTAB法提取6份菠菜品种的DNA,对AFLP反应体系中酶切连接、预扩、选扩等实验条件进行了优化分析。结果]研究结果如下:(1)在PCR仪中酶切的效果比水浴的要好,酶切反应对酶切时间和DNA的浓度要求不敏感。(2)菠菜AFLP预扩选扩体系的反应体积为20μl,Mg2+(25mmol/L)1.2μl,dNTPs(2.5mmol/L)1.6μl,Taq-polymerase(5U/μl)0.2μl最佳。结论]该研究建立的AFLP反应体系对与菠菜基因组分析时稳定有效的,该技术体系为菠菜品种亲缘关系的AFLP分子标记分析和遗传育种等提供一定的理论基础。

A Preliminary Study on the Establishment of AFLP Reaction System in Spinach

Objective] This study aimed to establish an AFLP reaction system for spinach. Method] Modified CTAB method was used for DNA extraction from six varieties of spinach. Experimental conditions in enzyme digestion and ligation, pre-amplification and selective amplification in the AFLP reaction system were analyzed and optimized. Result] The results showed that: (1) enzyme digestion had better effects in the PCR instrument than in a water bath and it was not sensitive to the digestion time and DNA concentrations; (2) a total volume of 20 μl, containing 1.2 μl of Mg2+ (25 mmol/L), 1.6 μl of dNTPs (2.5 mmol/L) and 0.2 μl of Taq-polymerase (5 U/μl) was the optimal condition for pre-amplification and selective amplification reactions. Conclusion] The AFLP system established in this study was stable and efficient, which provided a theoretical basis for AFLP molecular marker analysis of genetic relationship between spinach varieties and genetic breeding.