利用正交试验优化玫瑰SRAP-PCR反应体系

Optimization for SRAP-PCR System of Rosa rugosa Based on Orthogonal Design

采用L16(45)正交试验对玫瑰SRAP-PCR反应体系进行优化。结果表明,各因素对PCR反应的影响程度从大到小依次为:Taq酶,dNTPs,引物,Mg2+,模板;建立了玫瑰SRAP-PCR反应最佳体系(25μL)为Mg2+2.0mmol/L,dNTPs0.20mmol/L,Taq酶1.5U,引物0.25μmol/L,模板1.0ng/μL;采用不同的模板和引物对体系进行验证,表明该体系适合于玫瑰的SRAP-PCR反应。

The orthogonal design was employed to optimize SRAP-PCR amplification. The results showed that the order of factors which affect on the result of PCR were Taq polymerase, primers, dNTPs, Mg2＋ and DNA. A suitable SRAP-PCR system for Rose rugosa was that total 25 μL reaction system containing 2.0 mmol/L Mg2＋, 0.20 mmol/L dNTPs, 1.5U Taq polymerase, 0.25μmol/L primers and 1.0 ng/μL DNA. Use other primers and DNAs to test the system, the results showed that the system can be used in Rose rugosa.