仁用杏SRAP-PCR体系的正交设计优化

改良CTAB法提取仁用杏基因组DNA,用于建立SRAP-PCR体系。采用L16（44）正交设计试验,对影响仁用杏SRAP-PCR反应体系的4因素（dNTPs浓度、Mg^2＋浓度、引物浓度、Taq聚合酶量）4水平进行筛选,PCR结果经统计软件SPSS V13.0分析,并对退火温度进行了摸索,确立了仁用杏SRAP-PCR的优化体系：总体积为20 L,dNTPs浓度为0.2 mmol/L,Mg^2＋浓度为2.5mmol/L,引物浓度为0.9μmol/L,Taq聚合酶量为1.5 U,退火温度为52℃。本优化体系为仁用杏资源遗传分析奠定了技术基础。

Optimization of SRAP-PCR system for kernelled apricot by orthogonal design

The top-quality genomic DNA extracted from leaves of kernelled apricot with an improved method of CTAB was applied to optimize the SRAP-PCR system.The four levels of four factors(dNTPs,Mg2+,primers,Taq DNA polymerase) in SRAP-PCR system were studied by L16(44) orthogonal design.The electrophoresis profiles of SRAP-PCR were analyzed by software SPSS V13.0.The annealing temperature of PCR reaction was also optimized.The optimum SRAP-PCR system was shown as follows: 0.2 mmol/L dNTPs,2.5 mmol/L Mg2+,0.9 μmol/L primers,1.5 U Taq DNA polymerase in reaction solution of 20μL and annealing temperature at 52 ℃.The optimum SRAP-PCR system provides some technical foundations for the study of the phylogenetic relationship of kernelled apricot.