摘要
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.
出处
《草原与草坪》
CAS
2007年第3期25-27,共3页
Grassland and Turf
基金
甘肃省333人才工程项目
甘肃农业大学创新基金(GAU-CX0511)
关键词
抗蚜苜蓿
SSR
体系优化
aphid-resistant alfalfa
SSR
system optimization