摘要
采用正交设计方法,对影响番石榴SRAP反应体系的Mg2+、dNTPs、引物、TaqDNA聚合酶和模板DNA浓度等进行了优化,建立了适用于番石榴的SRAP反应体系。该优化的20μL反应体系中包含2.5mmol/LMg2+,0.15mmol/LdNTPs,0.4μmol/L引物,1.5UTaqDNA聚合酶和20ng模板DNA。利用该优化体系通过64对SRAP引物组合对5份番石榴材料进行了SRAP-PCR扩增,结果表明SRAP引物及优化后的反应体系能够有效地用于番石榴种质资源鉴定及遗传多样性分析等研究。
Orthogonal design was applied to optimize SRAP-PCR system of Guava（Psidium guajava L.）.Five main factors influencing on SRAP-PCR,including Mg 2 ＋,dNTPs,primer,Taq DNA polymerase and DNA template were examined by using orthogonal design.The results showed that the optimum concentration was Mg 2＋ 2.5 mmol/L,dNTPs 0.15 mmol/L,primer 0.4 μmol/L,Taq DNA polymerase 1.5 U,20 ng template DNA with a total volume of 20 μL reaction solution,which suitable for P.guajava SRAP-PCR amplification.The molecular identification of 5 varieties of P.guajava was conducted by 64 SRAP primer combinations with the optimal SRAP marker system.It showed that the SRAP primers and its optimized system could be applied in the germplasm identification and genetic diversity analysis of P.guajava effectively.
出处
《中国农学通报》
CSCD
北大核心
2011年第22期219-223,共5页
Chinese Agricultural Science Bulletin
基金
广西壮族自治区直属公益性科研院所基本科研业务费专项资金项目(桂热研201006)
关键词
番石榴
SRAP
正交设计
体系优化
guava（P.guajava L.）
SRAP
orthogonal design
optimization
