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辣木SCoT-PCR反应体系建立和引物筛选
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  • 英文篇名:Establishment of SCoT-PCR Reaction System and Primer Selection for Moringa oleifera Lam
  • 作者:林艺华 ; 郑涛 ; 蔡坤秀 ; 林宗铿
  • 英文作者:Lin Yihua;Zheng Tao;Cai Kunxiu;Lin Zongkeng;Fujian Institute of Tropical Crops;
  • 关键词:辣木 ; SCoT ; 反应体系 ; 引物筛选
  • 英文关键词:Moringa oleifera;;SCoT;;Reaction system;;Primer selection
  • 中文刊名:FZZW
  • 英文刊名:Molecular Plant Breeding
  • 机构:福建省热带作物科学研究所;
  • 出版日期:2018-07-14
  • 出版单位:分子植物育种
  • 年:2018
  • 期:v.16
  • 基金:福建省公益类科研院所基本科研专项(2016R1027-1);; 福建省自然科学基金项目(2016J01141)共同资助
  • 语种:中文;
  • 页:FZZW201813040
  • 页数:6
  • CN:13
  • ISSN:46-1068/S
  • 分类号:225-230
摘要
SCoT-PCR是一种单引物扩增分子标记方法,具有丰富的多态性,低廉的成本和较好的引物通用性,已在多种植物上加以应用。本研究采用L16(45)正交试验设计,对SCoT-PCR的影响因素进行优化试验,建立了适用于辣木的最佳SCoT-PCR反应体系,并对40条SCoT引物进行筛选。结果表明,25μL最佳反应总体积包含2.0 mmol/L Mg~(2+),0.3 mmol/L d NTPs,1.5 U ExTaq,1.25μmol/L引物,50 ng DNA模板,其中,dNTPs对该体系的影响程度最大。此外,40条SCoT引物共筛选出15条多态性好且适合辣木扩增的引物。该体系的建立为今后利用SCoT分子标记分析辣木种质资源鉴定与多样性、构建指纹图谱和辅助选择育种提供新的技术手段。
        SCoT-PCR is a molecular marker method of single primer amplification and is widely used in various plants for its abundant polymorphism, cheap cost and primer universality. The study used the L16(45) orthogonal design to carry out the optimizing experiment of the influence factors of SCoT-PCR. We established the optimum SCoT-PCR reaction system for Moringa oleifera and screened 40 SCoT primers. The result showed that the total volume of the25 μL best reaction contained 2.0 mmol/L Mg2+, 0.3 mmol/L d NTPs, 1.5 U Ex Taq, 1.25 μmol/L primer and 50 ng DNA template, among which d NTPs had the greatest impact on the system. Furthermore, 15 primers with good polymorphism which were fit for Moringa amplification were screened from the 40 SCoT primers. The establishment of this system might provide new technological means for future use of SCoT molecular markers to analyze the identification and diversity of Moringa oleifera germplasm resources and to build fingerprint and marker assisted selection.
引文
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