番木瓜SCoT反应体系建立及引物筛选

Establishment of SCoT Amplification System and Primer Selection for Carica papaya

摘要:

采用单因素试验方法对番木瓜(Carica papaya L.) SCoT-PCR反应体系进行了优化, 并对引物进行了筛选。结果表明, 在20 μL的番木瓜SCoT-PCR反应体系中, 包含2.0 mmol L^-1 Mg²⁺, 0.3 mmol L^-1 dNTPs, 0.8 μmol L^-1引物, 30 mg L^-1模板DNA和1.0 U Taq聚合酶。运用该体系对不同引物和22个番木瓜品种进行验证, 扩增的稳定性、可靠性良好, 且分辨率较高, 同时筛选出多态性丰富的5条引物和5对引物组合。因此, 优化的番木瓜SCoT-PCR反应体系可用于种质鉴定和遗传多样性分析。

Abstract:

The SCoT-PCR system for Carica papaya was optimized by using single factor experiment method, such as Mg²⁺, dNTPs, primer, Taq polymerase and DNA template, and the SCoT primers were selected with optimized system. The results showed that the optimum concentrations for Carica papaya SCoT-PCR amplification were 2.0 mmol L^-1 Mg²⁺, 0.3 mmol L^-1 dNTPs, 0.8 μmol L^-1 primer, 1.0 U Taq polymerase, and 30 mg L^-1 template DNA in total 20 μL reaction system, respectively. The optimum SCoT-PCR system was tested on 22 varieties of Carica papaya by different SCoT primers, obtaining steady, reliable and high resolution bands. Five primers and five pairs of primer combination were selected with abundant polymorphism. It was suggested that the optimum SCoT-PCR system and polymorphism primers could be used in the germplasm identification and genetic diversity analysis of Carica papaya.