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为建立最佳的宫粉紫荆SRAP-PCR反应体系,采用单因素和L16(45)正交试验设计对反应体系中的模板DNA、Mg2+、引物浓度、d NTPs和Taq聚合酶进行优化。表明宫粉紫荆SRAP-PCR 25μL反应体系的最佳组合为:模板DNA 50 ng、Mg2+2.25 mmol/L、引物0.25μmol/L、d NTPs 0.30 mmol/L、Taq酶1.5 U。并利用优化的SRAP-PCR体系进行验证,表明不同的宫粉紫荆样本均能扩增出清晰且带型基本一致的谱带,表明本试验建立的SRAP-PCR体系稳定,可用于今后开展宫粉紫荆种质资源遗传多样性研究、品种鉴定、优良品种筛选和近缘种杂交育种等研究工作。
In order to establish the optimal SRAP-PCR reaction system for Bauhinia rubra, template DNA, Mg2 +, primer concentration, dNTPs and Taq polymerase in the reaction system were optimized by single factor and L16 (45) orthogonal design. The results showed that the best combination of SRAP-PCR 25μL reaction system was: template DNA 50ng, Mg2 + 2.25mmol / L, primer 0.25μmol / L, d NTPs 0.30mmol / L, Taq enzyme 1.5U. The optimized SRAP-PCR system was used to verify that different samples of Bauhinia virginiana could amplify clear and band-like bands, indicating that the SRAP-PCR system established in this experiment is stable and can be used for the future development of uterine powder Bauhinia germplasm resources genetic diversity research, identification of varieties, selection of fine varieties and related species of cross breeding and other research work.