Optimization of AFLP reaction system in recessive-male sterile Sesamum indicum L. | 隐性核不育芝麻AFLP反应体系的优化
2011
Yang Minmin, Chinese Academy of Agricultural Sciences, Wuhan(China), Oil Crops Research Institute | Liu Hongyan, Chinese Academy of Agricultural Sciences, Wuhan(China), Oil Crops Research Institute | Zuo Yang, Chinese Academy of Agricultural Sciences, Wuhan(China), Oil Crops Research Institute
Chinese. 摘 要：以芝麻细胞核雄性不育系95ms-5为材料,对AFLP反应体系中的酶切连接、预扩增和选择性扩增中的关键性因素进行了优化。结果表明,在模板DNA质量浓度为200ng/μL、37℃酶切连接6h的情况下,预扩增中最佳Mg2＋浓度为1.0mmol/L,dNTP最佳浓度为0.3mmol/L,Taq酶量以0.5U为宜,预扩增引物终浓度为0.4mmol/L;选择性扩增中,预扩增产物稀释10倍后,以Mg2＋0.8mmol/L、dNTP 0.2mmol/L、Taq酶1.0U、选扩引物0.6mmol/L为宜。
Show more [+] Less [-]English. Key factors affecting the digestion-ligation,pre-amplification and selective amplification in amplified fragment length polymorphism（AFLP） analysis in sesame（Sesamum indicum L.） were optimized.Results showed that the best protocol was 200 ng/μL template DNA,digestion-ligation at 37 ℃ for 6 h,1.0 mmol/L Mg2＋,0.3 mmol/L dNTP,0.5 U Taq DNA polymerase in the pre-amplification system and pre-amplification products diluted by 10 times,1.0 mmol/L Mg2＋,0.2 mmol/L dNTP,and 1.0 U Taq DNA polymerase,0.6 mmol/L primers in the selective amplification.
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