橡胶树SRAP体系优化及其PCR引物组合的筛选  

Optimization of SRAP System and Selection of PCR Primer Combinations in Hevea brasiliensis

作　　者：王惠君[1] 王文泉[2] 卢诚[2] 王海燕[2] 陈新[2] 

机构地区：[1]琼州学院,海南三亚572022 [2]中国热带农业科学院/热带生物技术研究所,海南海口571101

出　　处：《贵州农业科学》2015年第2期29-34,共6页Guizhou Agricultural Sciences

基　　金：国家自然科学基金项目"木薯SNP遗传图谱构建与抗寒性QTL精细定位"(31171230);中央级科研院所基本科研业务费项目"麻疯树种质资源的收集与遗传改良"(ITBBZX0843)

摘　　要：为橡胶树遗传多样性分析及遗传图谱的构建奠定基础,分析引物浓度、Mg2+浓度、dNTPs浓度、模板DNA用量、Taq酶用量等因素对SRAP-PCR扩增反应的影响。结果表明,橡胶树SRAP-PCR的适宜扩增反应体系(20μL)为:引物0.2μmol/L,Mg2+2.0mmol/L,dNTPs 0.20mmol/L,模板DNA50ng,Taq酶0.5U。利用该体系从400对引物组合中初筛出28组多态性好的引物组合,获得250条清晰带,多态性比率为62.8%。To lay the foundation for evaluation of genetic diversity analysis and genetic linkage map in H.brasiliensis,factors influencing SRAP-PCR amplification system were analyzed,including primer concentration,Mg2＋ concentration,dNTPs concentration,DNA template dosage and Taq polymerase. Results：A suitable SRAP-PCR analysis reaction system for H.brasiliensis was obtained.The reaction mixture （20 μL）contained primer 0.2 μmol/L,Mg2＋ 2.0 mmol/L,dNTPs 0.20 mmol/L,50 ng of genomic DNA template,0.5 U of Taq polymerase.Conclusion：using optimized SRAP-PCR system,28 pairs of primers were screened out of 400 pairs of primers. 250 clear bands were gained and the polymorphism rate was 62.8%.

关 键 词：橡胶树 SRAP 体系优化 引物组合筛选 

分 类 号：S722.3[农业科学—林木遗传育种]