CAJ | 학술논문

采用正交设计,从Mg~(2+)、dNTPs、引物三种因素3个水平对青花菜SRAP反应体系进行优化,确立适合青花菜的SRAP反应体系,并在6个青花菜和花椰菜品种中进行验证.结果表明:20 μL反应体系中适宜体系的浓度dNTPs 为0.2 mmol/L、Mg~(2+)为1.25 mmol/L、上下引物各0.4 μmol/L、DNA为20 ng、Taq酶1.2 U、退火温度为50 ℃.
채용정교설계,종Mg~(2+)、dNTPs、인물삼충인소3개수평대청화채SRAP반응체계진행우화,학립괄합청화채적SRAP반응체계,병재6개청화채화화야채품충중진행험증.결과표명:20 μL반응체계중괄의체계적농도dNTPs 위0.2 mmol/L、Mg~(2+)위1.25 mmol/L、상하인물각0.4 μmol/L、DNA위20 ng、Taq매1.2 U、퇴화온도위50 ℃.
By an orthogonal design the reaction system of brocco's SRAP(sequence-related amplified polymorphism)was optimized in terms of Mg~(2+),dNTPs and primers three factors and their three levels,and then the optimized SRAP reaction system was verified with 6 broccoli and cauliflower varieties.The result indicated that in a 20 μL reaction system the appropriate concentrations of dNTPs,Mg~(2+),and forward and reverse primers were respectively 0.2 mmol/L,1.25 mmol/L,0.4 μmol/L and 0.4 μmol/L,the DNA of template was 20 ng,and Taq DNA polymerase 1.2 U,the best annealing temperature was 50 ℃.The results offer the technique basis of using SRAP technology to study broccoli germplasm resource.