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%.
出处
《贵州农业科学》
CAS
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)
关键词
橡胶树
SRAP
体系优化
引物组合筛选
Hevea brasiliensis
SRAP
system optimization
primer combinations screening
