能源植物芒的SRAP分子标记体系建立与优化

Optimization of SRAP-PCR System in Miscanthus sinensis

以芒总DNA为材料,利用单因素分析法对影响SRAP反应体系的Mg2+、dNTPs、TaqDNA聚合酶等3个因素进行了优化。研究结果表明:最佳的10μL反应体系为1μL10×TaqBuffer、DNA20ng、Mg2+2mmol/L、dNTPs0.5mmol/L、TaqDNA聚合酶0.6U、正反向Primer浓度均为0.8μmol/L。SRAP-PCR反应体系的建立和优化,为今后利用SRAP标记技术开展芒的遗传多样性研究和分子标记辅助选择育种研究提供了一个技术支持。

For optimizing SRAP-PCR reaction system of Miscanthus sinensis,the single factor experiment included three factors,such as Taq DNA polymerse,dNTPs and Mg 2 ＋,were optimized by using the total genomic DNA in this research.In conclusion the reaction system obtained were as followed：1 μL 10 × Taq Buffer,0.5 mmol/L dNTPs,0.6 U Taq DNA polymerase,2 mmol/L MgCl 2,20 ng DNA,0.8 μmol/L primer and with total 10 μL reaction solution.The results provided a useful technique for the genetic variation research and molecular marker-assisted breeding of M.sinensis.