不结球白菜SRAP反应体系的建立与优化

The Optimization of SRAP Amplification Program and System in Non-heading Chinese Cabbage

以菜薹基因组DNA为模板,通过正交试验设计,从Mg2+、Taq酶、dNTPs、引物及模板5种因素4个水平对不结球白菜SRAP反应体系进行优化,建立了适合不结球白菜的SRAP-PCR优化反应体系。利用该优化体系,选用30对芸薹属SRAP引物,对5份不结球白菜的基因组进行扩增,筛选出7对具有2条以上特异扩增条带的SRAP引物,验证了该优化体系的稳定性。

Using flowering Chinese cabbage （Brassica parachinensis L. H. Bariley） genome DNA as template,the major components of SRAP including concentrations of Mg2 ＋ ,dNTPs,Taq DNA polymerase,primers and template DNA were optimized in this study by orthogonal design with five factors and four levels respectively. The optimum SRAP-PCR reaction system suit for non-heading Chinese cabbage was established. Using this system,five accessions of non-heading Chinese cabbages were used as template DNA to be amplified by 30 SRAP primer pairs designed from Brassica genus. Seven SRAP primer pairs revealed two bands or more in 5 non-heading Chinese cabbage cultivars were selected. The result shows the optimization of SRAP System in non-heading Chinese Cabbage in stable.