东部白松SRAP反应体系的建立和优化

Establishment and Optimization of SRAP Reaction System in Pinus strobus

以东部白松针叶DNA为模板,采取正交实验设计L16(45)对SRAP-PCR反应体系的5个因素(Taq酶,Mg2+,dNTPs,模板DNA,引物)在4个水平上进行优化试验。结果表明:确定东部白松SRAP-PCR最佳反应体系(20μL):Taq酶0.5 U,Mg2+1.5 mmol/L,dNTPs 0.15mmol/L,模板DNA 50 ng,引物0.1μmol/L。

To construct the optimum SRAP reaction system of Pinus strobus,Pinus strobus needles DNA was abstracted as template and orthogonal experimental design L16（45） was used.And the comprehensive effect of four levels of five factors（Taq polymerase,Mg2＋,dNTPs,DNA template and primer） was assessed.,The results showed that an optimal and stable SRAP-PCR system for Pinus Strobus was obtained：0.5 U Taq polymerase,1.5 mmol/L Mg2＋,0.15 mmol/L dNTPs,50 ng DNA template,0.1 μmol/L primer,within total 20 μL reaction solution.