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目标区域扩增多态性(target region amplified polymorphism,TRAP)是通过一个依据EST序列设计的固定引物和一个针对外显子或内含子特点设计的随机引物对开放阅读框(open reading frames,ORFs)进行扩增。本研究首次建立了适于花生TRAP分析的PCR扩增体系:在15μl总反应体系中,模板DNA50ng,引物浓度1.0μmol/L,dNTPs浓度0.25mmol/L,Taq DNA聚合酶1U。利用该体系对24个花生品种DNA进行扩增,得到了清晰稳定的条带,且这些条带具有一定的多态性,说明所建立的体系可用于花生遗传多样性分析。
Target region amplified polymorphism (TRAP) is achieved by using a pair of open reading frames (ORFs) based on a set of fixed primers designed according to the EST sequence and a random primer designed for exon or intron features ). In this study, a PCR amplification system suitable for TRAP analysis of peanut was established for the first time. In a total reaction volume of 15μl, 50ng template DNA, 1.0μmol / L primer, 0.25mmol / L dNTPs and 1U Taq DNA polymerase. Using this system, we amplified the DNA of 24 peanut cultivars and obtained clear and stable bands. These bands have some polymorphism, indicating that the established system can be used for genetic analysis of peanut genetic diversity.