抗蚜紫花苜蓿品系SSR体系的建立

Optimization of SSR technique system on aphid-resistant alfalfa

以抗蚜紫花苜蓿品系HA-3为材料,研究了SSR反应体系中各个影响因素的浓度和用量,确定了适合苜蓿抗蚜基因定位的SSR体系:TaqDNA聚合酶0.2μL(5 U/μL),10×PCR Buffer(Mg2+Plus)2.5μL,dNTP Mixture 2μL(各2.5 mmol/L),引物1.5μL(10μmol/L),模板DNA 1μL,ddH2O16.3μL补足25μL。利用该体系进行扩增,所得谱带清晰、稳定、非特异带少。

An aphid-resistant alfalfa line （HA-3） was used to investigate proper concentrations of different reagents in SSR technique system and to seek the best combination of factors. The proper SSR technique system was established and the proper concentrations were as follows. Taq DNA polymerase was 0. 2 L（5 U/ L）, 10 PCR buffer （Mg^2＋ Plus） was 2. 5 L,dNTP mixture was 2 μL （2. 5 retool/L）, the primer was 1.5 μL （10 μmol/L）,the template DNA was 1μL, the ddH20 was 16.3btL, the total volume was 25μL.. Amplification bands were clear, standard and non-especial bands were few by using this system.