LIU Wei,CHEN Gang,YAN Ting-wu,et al.Optimization of SRAP Reaction System in Acer rubrum[J].Northern Horticulture,2015,39(02):79-83.[doi:10.11937/bfyy.201502023]
美国红枫SRAP反应体系优化
- Title:
- Optimization of SRAP Reaction System in Acer rubrum
- Keywords:
- Acer rubrum; SRAP; system optimization
- 文献标志码:
- A
- 摘要:
- 以美国红枫基因组DNA为模板,采取L16(45)正交实验设计,对影响扩增反应的5个主要因素进行优化。结果表明:在20μL反应体系中,各因素的最佳配比为Mg2+浓度1.5 mmol/L,dNTPs浓度2.5 mmol/L,引物浓度1.5 μmol/L,模板DNA用量 50 ng,Taq DNA聚合酶用量0.5 U。该体系的建立为SRAP技术在美国红枫分子辅助育种实践中提供了技术基础。
- Abstract:
- Using the genomic DNA of Acer rubrum as template,L16 (45) orthogonal experiment was designed to optimized the 5 major factors influencing the amplification reaction.The results showed that the optimum ratio of each factor in 20μL reaction system was 1.5 mmol/L of Mg2+ concentration,2.5 mmol/L of dNTPs concentration,1.5 μmol/L of primer concentration,50 ng of template DNA,and 0.5 U of Taq DNA polymerase.The establishment of this system provided a technological basis for the application of SRAP in molecular assisted breeding in Acer rubrum.
参考文献/References:
[1]Li G,Quiros C F.Sequence-related amplified polymorphism(SRAP),a new marker system based on a simple PCR reaction:its application to mapping and gene tagging in Brassica[J].Theor Appl Genel,2001,103:455-461.
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备注/Memo
第一作者简介:刘巍(1988-),男,硕士研究生,研究方向为生物化学与分子生物学。责任作者:陈罡(1980-),男,硕士,高级工程师,现主要从事林木分子遗传育种等研究工作。E-mail:chengang1625@163.com.基金项目:国家林业局“948”资助项目(2011-4-25);辽宁省科技计划资助项目(201404214)。